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  • 95
    Millipore lapatinib
    The combination of Notch and ErbB1/2 receptor inhibition reduces acinar size and mammosphere formation regardless of ErbB2 status. MCF10DCIS.com and SUM225 cells were treated with control, DAPT, <t>lapatinib</t> or combination (comb) in (A) matrigel culture and (B) mammosphere culture. (A) Cells were treated from day 0 and media/inhibitor were changed every 3 days. After 21 days in culture the sizes of the acini were measured ( µm).(B) Mammosphere-forming efficiency (MFE) was calculated by dividing the number of mammospheres formed by the original number of cells seeded and is expressed as a percentage compared to control. Mean±standard error of 3 independent experiments, Man Witney U test, two-tailed, NSD – No significant difference.
    Lapatinib, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 135 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Selleck Chemicals lapatinib
    Clinical characteristics and spectrum of ERBB2 mutations in breast cancer patients and construction of ERBB2 cell lines. (A) Left: follow-up computed tomography (CT) image taken 2 months after initiation of <t>lapatinib</t> treatment, with significant pulmonary metastasis in multiple regions after two cycles of lapatinib treatment. Right: follow-up CT images taken at 4 months after initiation of lapatinib treatment, with more significant pleural effusion. (B) The gene sequencing result of the tumor after 4-month treatment with lapatinib (whole blood sample by means of ctDNA detection). The insertion mutation (c.2339_2340-ins-CGGCTCCCC, p.P780_Y781insGSP) was found in ERBB2 exon 20. The diagram of insertion mutation of ERBB2. Wild type, WT; P780-Y781 mutation, MT. (C) Three groups of lentiviruses were added to the MDA-MB-231 or MCF-7 culture to the validate the transfection efficacy (through observing the fluorescence under a fluorescence microscope). (D) The expression level of ERBB2-WT and ERBB2-MT were determined by western blot.
    Lapatinib, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 99/100, based on 512 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lapatinib/product/Selleck Chemicals
    Average 99 stars, based on 512 article reviews
    Price from $9.99 to $1999.99
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    91
    BioVision lapatinib
    The inhibitory effect of <t>lapatinib</t> on the proliferation of 22 cancer cell lines and the rescue effect of fibroblast culture supernatant against lapatinib-induced inhibition. Lapanitib (1 μM) inhibits cell proliferation by more than 50% in the majority of the ESCC cell lines. Addition of fibroblast supernatant rescues growth inhibition by lapatinib and recovers cell proliferation by 20% or more in 10 cell lines.
    Lapatinib, supplied by BioVision, used in various techniques. Bioz Stars score: 91/100, based on 86 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lapatinib/product/BioVision
    Average 91 stars, based on 86 article reviews
    Price from $9.99 to $1999.99
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    93
    Selleck Chemicals lapatinib gw572016 ditosylate
    The inhibitory effect of <t>lapatinib</t> on the proliferation of 22 cancer cell lines and the rescue effect of fibroblast culture supernatant against lapatinib-induced inhibition. Lapanitib (1 μM) inhibits cell proliferation by more than 50% in the majority of the ESCC cell lines. Addition of fibroblast supernatant rescues growth inhibition by lapatinib and recovers cell proliferation by 20% or more in 10 cell lines.
    Lapatinib Gw572016 Ditosylate, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lapatinib gw572016 ditosylate/product/Selleck Chemicals
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    88
    Glaxo Smith lapatinib tykerb
    BT474 <t>lapatinib</t> resistant cells with prolonged treatment reactivate HER receptor activity . (A) Growth curves of UACC-812 and BT474 late stage lapatinib resistant cells (LLR) treated with different targeted therapies for nine days: trastuzumab (T) (10 μg/ml), lapatinib (L) (1 μM), trastuzumab plus lapatinib, or endocrine therapy, fulvestrant (F) (10 -7 M). Significance between groups was determined by multiple comparisons using the Sidak method (* P = 0.0008, BT474 LLR + L versus LLR + L + F, * P = 0.0044, BT474 LLR + L versus LLR + L + T; * P
    Lapatinib Tykerb, supplied by Glaxo Smith, used in various techniques. Bioz Stars score: 88/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lapatinib tykerb/product/Glaxo Smith
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    90
    Glaxo Smith lapatinib ditosylate tykerb tablets
    <t>Lapatinib</t> induces apoptosis in association with downregulation of CIP2A and p-Akt in TNBC cells A. Dose-escalation effects of lapatinib on CIP2A, p-Akt, and caspase 3 cleavage. Cells were exposed to lapatinib at the indicated doses for 48 hours. B. time-dependent analysis of CIP2A, p-Akt, and caspase 3 cleavage. Cells were exposed to lapatinib (10 μM) for 24, 36 and 48 hours. Cell lysates were prepared and assayed for these molecules by western blotting. Data are representative of three independent experiments. Apoptotic cells were determined by flow cytometry (sub-G1 analysis of propidium iodide -stained cells).
    Lapatinib Ditosylate Tykerb Tablets, supplied by Glaxo Smith, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Tocris lapatinib
    <t>Lapatinib</t> induces apoptosis in association with downregulation of CIP2A and p-Akt in TNBC cells A. Dose-escalation effects of lapatinib on CIP2A, p-Akt, and caspase 3 cleavage. Cells were exposed to lapatinib at the indicated doses for 48 hours. B. time-dependent analysis of CIP2A, p-Akt, and caspase 3 cleavage. Cells were exposed to lapatinib (10 μM) for 24, 36 and 48 hours. Cell lysates were prepared and assayed for these molecules by western blotting. Data are representative of three independent experiments. Apoptotic cells were determined by flow cytometry (sub-G1 analysis of propidium iodide -stained cells).
    Lapatinib, supplied by Tocris, used in various techniques. Bioz Stars score: 91/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Cell Signaling Technology Inc lapatinib
    PI3K/Akt signaling is required for VPA-induced upregulation of MICA and MICB in pancreatic cancer cells. (A, B) Quantitative real-time RT-PCR analysis. The VPA-induced upregulation of MICA and MICB mRNA expression were inhibited by the HER2/HER3 inhibitor <t>lapatinib</t> and the PI3K inhibitor LY294002, but not by the ATM/ATR inhibitor caffeine. Data are mean ± SD of a single experiment performed in triplicate, all results were reproducible in three independent experiments. * P
    Lapatinib, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 91/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lapatinib/product/Cell Signaling Technology Inc
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    91
    Novartis tykerb lapatinib east hanover
    PI3K/Akt signaling is required for VPA-induced upregulation of MICA and MICB in pancreatic cancer cells. (A, B) Quantitative real-time RT-PCR analysis. The VPA-induced upregulation of MICA and MICB mRNA expression were inhibited by the HER2/HER3 inhibitor <t>lapatinib</t> and the PI3K inhibitor LY294002, but not by the ATM/ATR inhibitor caffeine. Data are mean ± SD of a single experiment performed in triplicate, all results were reproducible in three independent experiments. * P
    Tykerb Lapatinib East Hanover, supplied by Novartis, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    The combination of Notch and ErbB1/2 receptor inhibition reduces acinar size and mammosphere formation regardless of ErbB2 status. MCF10DCIS.com and SUM225 cells were treated with control, DAPT, lapatinib or combination (comb) in (A) matrigel culture and (B) mammosphere culture. (A) Cells were treated from day 0 and media/inhibitor were changed every 3 days. After 21 days in culture the sizes of the acini were measured ( µm).(B) Mammosphere-forming efficiency (MFE) was calculated by dividing the number of mammospheres formed by the original number of cells seeded and is expressed as a percentage compared to control. Mean±standard error of 3 independent experiments, Man Witney U test, two-tailed, NSD – No significant difference.

    Journal: PLoS ONE

    Article Title: Combined Inhibition of ErbB1/2 and Notch Receptors Effectively Targets Breast Ductal Carcinoma In Situ (DCIS) Stem/Progenitor Cell Activity Regardless of ErbB2 Status

    doi: 10.1371/journal.pone.0056840

    Figure Lengend Snippet: The combination of Notch and ErbB1/2 receptor inhibition reduces acinar size and mammosphere formation regardless of ErbB2 status. MCF10DCIS.com and SUM225 cells were treated with control, DAPT, lapatinib or combination (comb) in (A) matrigel culture and (B) mammosphere culture. (A) Cells were treated from day 0 and media/inhibitor were changed every 3 days. After 21 days in culture the sizes of the acini were measured ( µm).(B) Mammosphere-forming efficiency (MFE) was calculated by dividing the number of mammospheres formed by the original number of cells seeded and is expressed as a percentage compared to control. Mean±standard error of 3 independent experiments, Man Witney U test, two-tailed, NSD – No significant difference.

    Article Snippet: Calbiochem) and lapatinib (a HER2/EGFR tyrosine kinase inhibitor) using DMSO control.

    Techniques: Inhibition, Two Tailed Test

    Notch and ErbB1/2 receptor inhibition reduces the size of DCIS cell line acini. MCF10DCIS.com and SUM225 cells were grown in matrigel culture in the presence or absence of DAPT 5 and 10 µM (A), Lapatinib 0.05 and 0.1 µM (B). Cells were treated from day 0 and media/inhibitor was changed every 3 days. After 21 days in culture the size of the acini were measured (µm). (C) Bright field images of DCIS acini grown in the presence or absence of DAPT 5 and 20 µM or Lapatinib 0.05-and 0.1 µM. Scale bar represent 100 µm, graphs represent mean±standard error of 3 independent experiments, Man Witney U test, two-tailed, * p≤0.032, ** p≤0.008, *** p≤0.0001.

    Journal: PLoS ONE

    Article Title: Combined Inhibition of ErbB1/2 and Notch Receptors Effectively Targets Breast Ductal Carcinoma In Situ (DCIS) Stem/Progenitor Cell Activity Regardless of ErbB2 Status

    doi: 10.1371/journal.pone.0056840

    Figure Lengend Snippet: Notch and ErbB1/2 receptor inhibition reduces the size of DCIS cell line acini. MCF10DCIS.com and SUM225 cells were grown in matrigel culture in the presence or absence of DAPT 5 and 10 µM (A), Lapatinib 0.05 and 0.1 µM (B). Cells were treated from day 0 and media/inhibitor was changed every 3 days. After 21 days in culture the size of the acini were measured (µm). (C) Bright field images of DCIS acini grown in the presence or absence of DAPT 5 and 20 µM or Lapatinib 0.05-and 0.1 µM. Scale bar represent 100 µm, graphs represent mean±standard error of 3 independent experiments, Man Witney U test, two-tailed, * p≤0.032, ** p≤0.008, *** p≤0.0001.

    Article Snippet: Calbiochem) and lapatinib (a HER2/EGFR tyrosine kinase inhibitor) using DMSO control.

    Techniques: Inhibition, Two Tailed Test

    Notch and ErbB1/2 receptor inhibition reduces mammosphere forming efficiency. MCF10DCIS.com and SUM-225 cells were treated with DAPT 1–10 µM (A) or Lapatinib 0.25–2.5 µM (B) in mammosphere culture. Mammosphere-forming efficiency (MFE) was calculated by dividing the number of mammospheres formed by the original number of cells seeded and is expressed as a percentage compared to control. Mean±standard error n = 3, Man Witney U test, two-tailed, * p

    Journal: PLoS ONE

    Article Title: Combined Inhibition of ErbB1/2 and Notch Receptors Effectively Targets Breast Ductal Carcinoma In Situ (DCIS) Stem/Progenitor Cell Activity Regardless of ErbB2 Status

    doi: 10.1371/journal.pone.0056840

    Figure Lengend Snippet: Notch and ErbB1/2 receptor inhibition reduces mammosphere forming efficiency. MCF10DCIS.com and SUM-225 cells were treated with DAPT 1–10 µM (A) or Lapatinib 0.25–2.5 µM (B) in mammosphere culture. Mammosphere-forming efficiency (MFE) was calculated by dividing the number of mammospheres formed by the original number of cells seeded and is expressed as a percentage compared to control. Mean±standard error n = 3, Man Witney U test, two-tailed, * p

    Article Snippet: Calbiochem) and lapatinib (a HER2/EGFR tyrosine kinase inhibitor) using DMSO control.

    Techniques: Inhibition, Two Tailed Test

    Downstream signalling in DCIS cell line mammospheres. Western blot analysis of downstream targets of Notch and ErbB1/2 receptor signalling in (A) MCF10DCIS.com and (B) SUM225 cells after treatment with DAPT or Lapatinib for 7 days in non-adherent mammosphere culture. Treatments were added at time zero. pAKT = phospho-AKT; AKT = total AKT; pMAPK = phospho-MAPK; MAPK = total MAPK; NICD = Notch1 intracellular domain. β-actin was used as a loading control.

    Journal: PLoS ONE

    Article Title: Combined Inhibition of ErbB1/2 and Notch Receptors Effectively Targets Breast Ductal Carcinoma In Situ (DCIS) Stem/Progenitor Cell Activity Regardless of ErbB2 Status

    doi: 10.1371/journal.pone.0056840

    Figure Lengend Snippet: Downstream signalling in DCIS cell line mammospheres. Western blot analysis of downstream targets of Notch and ErbB1/2 receptor signalling in (A) MCF10DCIS.com and (B) SUM225 cells after treatment with DAPT or Lapatinib for 7 days in non-adherent mammosphere culture. Treatments were added at time zero. pAKT = phospho-AKT; AKT = total AKT; pMAPK = phospho-MAPK; MAPK = total MAPK; NICD = Notch1 intracellular domain. β-actin was used as a loading control.

    Article Snippet: Calbiochem) and lapatinib (a HER2/EGFR tyrosine kinase inhibitor) using DMSO control.

    Techniques: Western Blot

    Clinical characteristics and spectrum of ERBB2 mutations in breast cancer patients and construction of ERBB2 cell lines. (A) Left: follow-up computed tomography (CT) image taken 2 months after initiation of lapatinib treatment, with significant pulmonary metastasis in multiple regions after two cycles of lapatinib treatment. Right: follow-up CT images taken at 4 months after initiation of lapatinib treatment, with more significant pleural effusion. (B) The gene sequencing result of the tumor after 4-month treatment with lapatinib (whole blood sample by means of ctDNA detection). The insertion mutation (c.2339_2340-ins-CGGCTCCCC, p.P780_Y781insGSP) was found in ERBB2 exon 20. The diagram of insertion mutation of ERBB2. Wild type, WT; P780-Y781 mutation, MT. (C) Three groups of lentiviruses were added to the MDA-MB-231 or MCF-7 culture to the validate the transfection efficacy (through observing the fluorescence under a fluorescence microscope). (D) The expression level of ERBB2-WT and ERBB2-MT were determined by western blot.

    Journal: Biology Open

    Article Title: An insertion mutation of ERBB2 enhances breast cancer cell growth and confers resistance to lapatinib through AKT signaling pathway

    doi: 10.1242/bio.047662

    Figure Lengend Snippet: Clinical characteristics and spectrum of ERBB2 mutations in breast cancer patients and construction of ERBB2 cell lines. (A) Left: follow-up computed tomography (CT) image taken 2 months after initiation of lapatinib treatment, with significant pulmonary metastasis in multiple regions after two cycles of lapatinib treatment. Right: follow-up CT images taken at 4 months after initiation of lapatinib treatment, with more significant pleural effusion. (B) The gene sequencing result of the tumor after 4-month treatment with lapatinib (whole blood sample by means of ctDNA detection). The insertion mutation (c.2339_2340-ins-CGGCTCCCC, p.P780_Y781insGSP) was found in ERBB2 exon 20. The diagram of insertion mutation of ERBB2. Wild type, WT; P780-Y781 mutation, MT. (C) Three groups of lentiviruses were added to the MDA-MB-231 or MCF-7 culture to the validate the transfection efficacy (through observing the fluorescence under a fluorescence microscope). (D) The expression level of ERBB2-WT and ERBB2-MT were determined by western blot.

    Article Snippet: Following attachment, cells were incubated with different concentrations of lapatinib (S2111, Selleckchem, Houston, TX, USA) and/or perifosine (1.5–100 μM APExBIO, Houston, TX, USA) and cultured for 48 h. CCK8 agent (Dojindo Molecular Technologies, Inc., Japan) was added, and after 2 h, the absorbance at 450 nm was measured using a microplate reader (Bio-Rad Laboratories, Hercules, CA, USA).

    Techniques: Computed Tomography, Sequencing, Mutagenesis, Multiple Displacement Amplification, Transfection, Fluorescence, Microscopy, Expressing, Western Blot

    Mutant ERBB2 confers a resistance to lapatinib and is related to AKT signaling pathway. (A) Overexpression of ERBB2-MT enhances the lapatinib resistance (at 12.5 and 25 μM in comparison with ERBB-WT, both P

    Journal: Biology Open

    Article Title: An insertion mutation of ERBB2 enhances breast cancer cell growth and confers resistance to lapatinib through AKT signaling pathway

    doi: 10.1242/bio.047662

    Figure Lengend Snippet: Mutant ERBB2 confers a resistance to lapatinib and is related to AKT signaling pathway. (A) Overexpression of ERBB2-MT enhances the lapatinib resistance (at 12.5 and 25 μM in comparison with ERBB-WT, both P

    Article Snippet: Following attachment, cells were incubated with different concentrations of lapatinib (S2111, Selleckchem, Houston, TX, USA) and/or perifosine (1.5–100 μM APExBIO, Houston, TX, USA) and cultured for 48 h. CCK8 agent (Dojindo Molecular Technologies, Inc., Japan) was added, and after 2 h, the absorbance at 450 nm was measured using a microplate reader (Bio-Rad Laboratories, Hercules, CA, USA).

    Techniques: Mutagenesis, Over Expression

    P780-Y781 ERBB2 enhances colony-formation ability and but does not impact cell apoptosis. (A) Mutant ERBB2 dramatically elevated the colony counts. Typical photos of colony staining of three groups under vehicle, lapatinib, perifosine and lapatinib+perifosine. (B) The colony counts in the ERBB2-NC and ERBB2-WT groups. ERBB2-MT dramatically elevated the colony counts in comparison with WT (t=7.01, P

    Journal: Biology Open

    Article Title: An insertion mutation of ERBB2 enhances breast cancer cell growth and confers resistance to lapatinib through AKT signaling pathway

    doi: 10.1242/bio.047662

    Figure Lengend Snippet: P780-Y781 ERBB2 enhances colony-formation ability and but does not impact cell apoptosis. (A) Mutant ERBB2 dramatically elevated the colony counts. Typical photos of colony staining of three groups under vehicle, lapatinib, perifosine and lapatinib+perifosine. (B) The colony counts in the ERBB2-NC and ERBB2-WT groups. ERBB2-MT dramatically elevated the colony counts in comparison with WT (t=7.01, P

    Article Snippet: Following attachment, cells were incubated with different concentrations of lapatinib (S2111, Selleckchem, Houston, TX, USA) and/or perifosine (1.5–100 μM APExBIO, Houston, TX, USA) and cultured for 48 h. CCK8 agent (Dojindo Molecular Technologies, Inc., Japan) was added, and after 2 h, the absorbance at 450 nm was measured using a microplate reader (Bio-Rad Laboratories, Hercules, CA, USA).

    Techniques: Mutagenesis, Staining

    ERBB2-MT group has an elevated proportion of (S+G2+M) stages and the ERBB2-MT tumor has a higher p-AKT level. (A) In cycle distribution, the ERBB2-MT group showed an elevated proportion of (S+G2+M) stages compared with ERBB2-WT when exposed to lapatinib (t=14.97, P

    Journal: Biology Open

    Article Title: An insertion mutation of ERBB2 enhances breast cancer cell growth and confers resistance to lapatinib through AKT signaling pathway

    doi: 10.1242/bio.047662

    Figure Lengend Snippet: ERBB2-MT group has an elevated proportion of (S+G2+M) stages and the ERBB2-MT tumor has a higher p-AKT level. (A) In cycle distribution, the ERBB2-MT group showed an elevated proportion of (S+G2+M) stages compared with ERBB2-WT when exposed to lapatinib (t=14.97, P

    Article Snippet: Following attachment, cells were incubated with different concentrations of lapatinib (S2111, Selleckchem, Houston, TX, USA) and/or perifosine (1.5–100 μM APExBIO, Houston, TX, USA) and cultured for 48 h. CCK8 agent (Dojindo Molecular Technologies, Inc., Japan) was added, and after 2 h, the absorbance at 450 nm was measured using a microplate reader (Bio-Rad Laboratories, Hercules, CA, USA).

    Techniques:

    Coamplification of miR-4728 in HER2 -amplified breast cancer causes ERα-mediated NOXA down-regulation and its inhibition sensitizes them to lapatinib, while its overexpression rescues lapatinib-treated HER2 -amplified breast cancer cells from apoptosis. ( A ) The indicated HER2 -amplified breast cancer cell lines were transfected with the pLV-hsa-miR-4728-3p locker plasmid (miR-4728 inhibitor) or control plasmid expressing a scrambled sequence (vector control), and the corresponding lysates were subjected to immunoblotting for NOXA, ERα, HER2, and phospho-ERK. GAPDH and β-ACTIN were used as loading controls. ( B ) The indicated breast cancer cell lines were infected with control vector or a mir-4728 overexpressing construct, and the corresponding lysates were subjected to Western blotting and probed for NOXA, ERα, HER2, and β-ACTIN (loading control). ( C ) MDA-MB-361 cells were transfected with the appropriate constructs, as in A , treated with no drug or 200 nM fulvestrant for 24 h and probed for the indicated proteins. β-ACTIN was used as a loading control. ( D ) MDA-MB-361 cells were transfected as in A and treated with no drug and with 1 μM of lapatinib for the indicated time points. The corresponding lysates were subjected to Western blotting and probed for the indicated proteins. β-ACTIN was used as a loading control. ( E ) BT-474 cells were transfected with the pLV-hsa-miR-4728-3p locker plasmid (miR-4728 inhibitor) or control plasmid expressing a scrambled sequence (vector control), treated with no drug or 1 μM lapatinib for 24 h. The corresponding lysates were subjected to Western blotting and probed for the indicated proteins. β-ACTIN was used as a loading control. BT-474 cells were also infected with control vector or a miR-4728–overexpressing construct, like in B , and treated with no drug or 1 μM lapatinib for 36 h. The corresponding lysates were subjected to Western blotting and probed for the indicated proteins. ( F ) Suggested model for NOXA regulation by the HER2 amplicon in breast cancer. miR-4728-3p is coamplified with its host gene ( HER2 ) and proceeds to silence ESR1 expression. ERα encoded by ESR1 functions as a transcriptional factor of NOXA; therefore, miR-4728 coamplification leads to down-regulation of NOXA (“No Rx”: No drug).

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Coamplification of miR-4728 protects HER2-amplified breast cancers from targeted therapy

    doi: 10.1073/pnas.1717820115

    Figure Lengend Snippet: Coamplification of miR-4728 in HER2 -amplified breast cancer causes ERα-mediated NOXA down-regulation and its inhibition sensitizes them to lapatinib, while its overexpression rescues lapatinib-treated HER2 -amplified breast cancer cells from apoptosis. ( A ) The indicated HER2 -amplified breast cancer cell lines were transfected with the pLV-hsa-miR-4728-3p locker plasmid (miR-4728 inhibitor) or control plasmid expressing a scrambled sequence (vector control), and the corresponding lysates were subjected to immunoblotting for NOXA, ERα, HER2, and phospho-ERK. GAPDH and β-ACTIN were used as loading controls. ( B ) The indicated breast cancer cell lines were infected with control vector or a mir-4728 overexpressing construct, and the corresponding lysates were subjected to Western blotting and probed for NOXA, ERα, HER2, and β-ACTIN (loading control). ( C ) MDA-MB-361 cells were transfected with the appropriate constructs, as in A , treated with no drug or 200 nM fulvestrant for 24 h and probed for the indicated proteins. β-ACTIN was used as a loading control. ( D ) MDA-MB-361 cells were transfected as in A and treated with no drug and with 1 μM of lapatinib for the indicated time points. The corresponding lysates were subjected to Western blotting and probed for the indicated proteins. β-ACTIN was used as a loading control. ( E ) BT-474 cells were transfected with the pLV-hsa-miR-4728-3p locker plasmid (miR-4728 inhibitor) or control plasmid expressing a scrambled sequence (vector control), treated with no drug or 1 μM lapatinib for 24 h. The corresponding lysates were subjected to Western blotting and probed for the indicated proteins. β-ACTIN was used as a loading control. BT-474 cells were also infected with control vector or a miR-4728–overexpressing construct, like in B , and treated with no drug or 1 μM lapatinib for 36 h. The corresponding lysates were subjected to Western blotting and probed for the indicated proteins. ( F ) Suggested model for NOXA regulation by the HER2 amplicon in breast cancer. miR-4728-3p is coamplified with its host gene ( HER2 ) and proceeds to silence ESR1 expression. ERα encoded by ESR1 functions as a transcriptional factor of NOXA; therefore, miR-4728 coamplification leads to down-regulation of NOXA (“No Rx”: No drug).

    Article Snippet: The following drugs were purchased: for in vitro and in vivo studies (S-63845; Chemietek), Lapatinib Ditosylate (Tykerb) for in vitro and in vivo studies (M1802; Abmole), and Fulvestrant (S1191; Selleckchem).

    Techniques: Amplification, Inhibition, Over Expression, Transfection, Plasmid Preparation, Expressing, Sequencing, Infection, Construct, Western Blot, Multiple Displacement Amplification

    Comparison of the autophagic flux induction in parental (OE19 P) and Lapatinib-resistant (OE19 LR) OE19 cells. ( a ) Relative cell viability assessed by alamarBlue ® assay of OE19 P and OE19 LR cells, treated with dimethyl sulfoxide (DMSO) alone or 120nM of Lapatinib ( n = 3). ( b ) Quantification of FACS analysis comparing OE19 P and OE19 LR transduced with a mCherry-EGFP-LC3B construct (same treatment as in a). As a control, autophagy blocked conditions (addition of 5µM VPS34-IN1) were included, ( n = 4). The error bars represent SD, statistical significance was determined by Mann–Whitney U test: * p ≤ 0.05, ** p ≤ 0.01.

    Journal: International Journal of Molecular Sciences

    Article Title: Her2-Targeted Therapy Induces Autophagy in Esophageal Adenocarcinoma Cells

    doi: 10.3390/ijms19103069

    Figure Lengend Snippet: Comparison of the autophagic flux induction in parental (OE19 P) and Lapatinib-resistant (OE19 LR) OE19 cells. ( a ) Relative cell viability assessed by alamarBlue ® assay of OE19 P and OE19 LR cells, treated with dimethyl sulfoxide (DMSO) alone or 120nM of Lapatinib ( n = 3). ( b ) Quantification of FACS analysis comparing OE19 P and OE19 LR transduced with a mCherry-EGFP-LC3B construct (same treatment as in a). As a control, autophagy blocked conditions (addition of 5µM VPS34-IN1) were included, ( n = 4). The error bars represent SD, statistical significance was determined by Mann–Whitney U test: * p ≤ 0.05, ** p ≤ 0.01.

    Article Snippet: Lapatinib (Lap) (Selleckchem, LubioScience Luzern, Switzerland, S2111) was reconstituted in dimethyl sulfoxide (DMSO) and stock solutions were stored at −80 °C.

    Techniques: Alamar Blue Assay, FACS, Transduction, Construct, MANN-WHITNEY

    Induction of autophagic flux in OE19 upon Lapatinib treatment. ( a ) LC3B flux was assessed comparing control and BafilomycinA (BafA)-treated (200 nM, 2 h) OE19 upon Lapatinib treatment (120 nM, 24 h). LC3 band intensities were quantified using ImageJ software. Total protein was used as a loading control, and Phospho-Her2 for Lapatinib treatment ( n = 3). ( b ) LC3B flux was calculated from data in ( a ) as follows: BafA + -BafA − values for each condition. ( c ) FACS analysis of mCherry-EGFP-LC3B-expressing OE19 cells upon induction or blockade of autophagy with indication of the chosen cut-off value for high respectively low autophagic flux. ( d ) Quantification of the FACS analysis showing % of cells with high autophagic flux ( n = 3). Cells were treated as in ( a ). The error bars represent SD, statistical significance was determined by Mann–Whitney U test: * p ≤ 0.05, ** p ≤ 0.01.

    Journal: International Journal of Molecular Sciences

    Article Title: Her2-Targeted Therapy Induces Autophagy in Esophageal Adenocarcinoma Cells

    doi: 10.3390/ijms19103069

    Figure Lengend Snippet: Induction of autophagic flux in OE19 upon Lapatinib treatment. ( a ) LC3B flux was assessed comparing control and BafilomycinA (BafA)-treated (200 nM, 2 h) OE19 upon Lapatinib treatment (120 nM, 24 h). LC3 band intensities were quantified using ImageJ software. Total protein was used as a loading control, and Phospho-Her2 for Lapatinib treatment ( n = 3). ( b ) LC3B flux was calculated from data in ( a ) as follows: BafA + -BafA − values for each condition. ( c ) FACS analysis of mCherry-EGFP-LC3B-expressing OE19 cells upon induction or blockade of autophagy with indication of the chosen cut-off value for high respectively low autophagic flux. ( d ) Quantification of the FACS analysis showing % of cells with high autophagic flux ( n = 3). Cells were treated as in ( a ). The error bars represent SD, statistical significance was determined by Mann–Whitney U test: * p ≤ 0.05, ** p ≤ 0.01.

    Article Snippet: Lapatinib (Lap) (Selleckchem, LubioScience Luzern, Switzerland, S2111) was reconstituted in dimethyl sulfoxide (DMSO) and stock solutions were stored at −80 °C.

    Techniques: Software, FACS, Expressing, MANN-WHITNEY

    Combined Her2 and autophagy inhibitor treatment ( a ) Relative cell viability of OE19 cells treated with Lapatinib (120 nM) and/or autophagy inhibition either VPS34 inhibitor (VPS34-IN1) (5 µM) or chloroquine (CQ) (25 µM) at days 0 and 3 of the alamarBlue ® experiment ( n ≥ 3). ( b ) Cell counts of OE19 P and OE19 LR cells treated as in ( a ) but for 48 h. Values were normalized to DMSO control treated cells ( n = 7) ( c ) Colony numbers after treatment as described in ( b ), reseeded (2500 cells/well) in six-well plates and incubation for 14 days without treatment ( n = 5). The error bars represent SD, statistical significance was determined by Mann–Whitney U test: ns = non significant p > 0.05, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.

    Journal: International Journal of Molecular Sciences

    Article Title: Her2-Targeted Therapy Induces Autophagy in Esophageal Adenocarcinoma Cells

    doi: 10.3390/ijms19103069

    Figure Lengend Snippet: Combined Her2 and autophagy inhibitor treatment ( a ) Relative cell viability of OE19 cells treated with Lapatinib (120 nM) and/or autophagy inhibition either VPS34 inhibitor (VPS34-IN1) (5 µM) or chloroquine (CQ) (25 µM) at days 0 and 3 of the alamarBlue ® experiment ( n ≥ 3). ( b ) Cell counts of OE19 P and OE19 LR cells treated as in ( a ) but for 48 h. Values were normalized to DMSO control treated cells ( n = 7) ( c ) Colony numbers after treatment as described in ( b ), reseeded (2500 cells/well) in six-well plates and incubation for 14 days without treatment ( n = 5). The error bars represent SD, statistical significance was determined by Mann–Whitney U test: ns = non significant p > 0.05, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.

    Article Snippet: Lapatinib (Lap) (Selleckchem, LubioScience Luzern, Switzerland, S2111) was reconstituted in dimethyl sulfoxide (DMSO) and stock solutions were stored at −80 °C.

    Techniques: Inhibition, Incubation, MANN-WHITNEY

    NH125 enhances lapatinib-induced apoptosis in NPC cells. a , b and c CNE-2 and HONE-1 cells were treated with lapatinib (0-5 μM) or DMSO control for 48 h in the presence or absence of 0.25 μM NH125. a Annexin V-APC/7-AAD double staining was performed to detect apoptotic activity. b Cleaved PARP was examined by Western blot analysis. GAPDH was used as a loading control. c Flow cytometry was used to analyse cleaved PARP levels. Results are displayed as histograms. Each bar represents the mean ± standard deviation. *, P

    Journal: BMC Cancer

    Article Title: Inhibition of eEF-2 kinase sensitizes human nasopharyngeal carcinoma cells to lapatinib-induced apoptosis through the Src and Erk pathways

    doi: 10.1186/s12885-016-2853-5

    Figure Lengend Snippet: NH125 enhances lapatinib-induced apoptosis in NPC cells. a , b and c CNE-2 and HONE-1 cells were treated with lapatinib (0-5 μM) or DMSO control for 48 h in the presence or absence of 0.25 μM NH125. a Annexin V-APC/7-AAD double staining was performed to detect apoptotic activity. b Cleaved PARP was examined by Western blot analysis. GAPDH was used as a loading control. c Flow cytometry was used to analyse cleaved PARP levels. Results are displayed as histograms. Each bar represents the mean ± standard deviation. *, P

    Article Snippet: Inhibitors Lapatinib and NH125 were purchased from Selleck Chemicals (HOU, TX, USA).

    Techniques: Double Staining, Activity Assay, Western Blot, Flow Cytometry, Cytometry, Standard Deviation

    Lapatinib and NH125 exert synergistic effects through the Src and Erk signalling pathways. a , b , c and d CNE-2 and HONE-1 cells were treated with lapatinib, NH125 or their combination for 48 h. a Cell viability was assessed by the CCK-8 assay. b Inhibition of proliferation was measured by the crystal violet assay. c IC 50 isobologram of the lapatinib and NH125 combination treatment. In the isobologram, a plot to the left under the line indicates that the combination is synergistic. d Cleaved PARP, phosphorylated AKT, phosphorylated ERK and phosphorylated Src levels were examined by Western blot analysis. GAPDH was used as a loading control

    Journal: BMC Cancer

    Article Title: Inhibition of eEF-2 kinase sensitizes human nasopharyngeal carcinoma cells to lapatinib-induced apoptosis through the Src and Erk pathways

    doi: 10.1186/s12885-016-2853-5

    Figure Lengend Snippet: Lapatinib and NH125 exert synergistic effects through the Src and Erk signalling pathways. a , b , c and d CNE-2 and HONE-1 cells were treated with lapatinib, NH125 or their combination for 48 h. a Cell viability was assessed by the CCK-8 assay. b Inhibition of proliferation was measured by the crystal violet assay. c IC 50 isobologram of the lapatinib and NH125 combination treatment. In the isobologram, a plot to the left under the line indicates that the combination is synergistic. d Cleaved PARP, phosphorylated AKT, phosphorylated ERK and phosphorylated Src levels were examined by Western blot analysis. GAPDH was used as a loading control

    Article Snippet: Inhibitors Lapatinib and NH125 were purchased from Selleck Chemicals (HOU, TX, USA).

    Techniques: CCK-8 Assay, Inhibition, Crystal Violet Assay, Western Blot

    Silencing of eEF-2 kinase expression by RNA interference augments lapatinib-induced apoptosis in NPC cells. a and b NPC cells were transfected with a non-targeting RNA (NT) or siRNA targeting eEF-2 kinase (eEF-2 K siRNA) followed by treatment with lapatinib or DMSO for 48 h. a Cell viability was assessed by the CCK-8 assay. b Annexin V-APC/7-AAD double staining was performed to detect apoptotic activity. Results are displayed as histograms. Each bar represents the mean ± standard deviation. *, P

    Journal: BMC Cancer

    Article Title: Inhibition of eEF-2 kinase sensitizes human nasopharyngeal carcinoma cells to lapatinib-induced apoptosis through the Src and Erk pathways

    doi: 10.1186/s12885-016-2853-5

    Figure Lengend Snippet: Silencing of eEF-2 kinase expression by RNA interference augments lapatinib-induced apoptosis in NPC cells. a and b NPC cells were transfected with a non-targeting RNA (NT) or siRNA targeting eEF-2 kinase (eEF-2 K siRNA) followed by treatment with lapatinib or DMSO for 48 h. a Cell viability was assessed by the CCK-8 assay. b Annexin V-APC/7-AAD double staining was performed to detect apoptotic activity. Results are displayed as histograms. Each bar represents the mean ± standard deviation. *, P

    Article Snippet: Inhibitors Lapatinib and NH125 were purchased from Selleck Chemicals (HOU, TX, USA).

    Techniques: Expressing, Transfection, CCK-8 Assay, Double Staining, Activity Assay, Standard Deviation

    NH125 sensitizes NPC cells to lapatinib. a , b and c NPC cells were treated with lapatinib or DMSO for 48 h in the presence or absence of 0.25 μM NH125. a Cell viability was assessed by the CCK-8 assay. Results are expressed as means ± standard deviation. *, P

    Journal: BMC Cancer

    Article Title: Inhibition of eEF-2 kinase sensitizes human nasopharyngeal carcinoma cells to lapatinib-induced apoptosis through the Src and Erk pathways

    doi: 10.1186/s12885-016-2853-5

    Figure Lengend Snippet: NH125 sensitizes NPC cells to lapatinib. a , b and c NPC cells were treated with lapatinib or DMSO for 48 h in the presence or absence of 0.25 μM NH125. a Cell viability was assessed by the CCK-8 assay. Results are expressed as means ± standard deviation. *, P

    Article Snippet: Inhibitors Lapatinib and NH125 were purchased from Selleck Chemicals (HOU, TX, USA).

    Techniques: CCK-8 Assay, Standard Deviation

    Comparison of the autophagic flux induction in parental (OE19 P) and Lapatinib-resistant (OE19 LR) OE19 cells. ( a ) Relative cell viability assessed by alamarBlue ® assay of OE19 P and OE19 LR cells, treated with dimethyl sulfoxide (DMSO) alone or 120nM of Lapatinib ( n = 3). ( b ) Quantification of FACS analysis comparing OE19 P and OE19 LR transduced with a mCherry-EGFP-LC3B construct (same treatment as in a). As a control, autophagy blocked conditions (addition of 5µM VPS34-IN1) were included, ( n = 4). The error bars represent SD, statistical significance was determined by Mann–Whitney U test: * p ≤ 0.05, ** p ≤ 0.01.

    Journal: International Journal of Molecular Sciences

    Article Title: Her2-Targeted Therapy Induces Autophagy in Esophageal Adenocarcinoma Cells

    doi: 10.3390/ijms19103069

    Figure Lengend Snippet: Comparison of the autophagic flux induction in parental (OE19 P) and Lapatinib-resistant (OE19 LR) OE19 cells. ( a ) Relative cell viability assessed by alamarBlue ® assay of OE19 P and OE19 LR cells, treated with dimethyl sulfoxide (DMSO) alone or 120nM of Lapatinib ( n = 3). ( b ) Quantification of FACS analysis comparing OE19 P and OE19 LR transduced with a mCherry-EGFP-LC3B construct (same treatment as in a). As a control, autophagy blocked conditions (addition of 5µM VPS34-IN1) were included, ( n = 4). The error bars represent SD, statistical significance was determined by Mann–Whitney U test: * p ≤ 0.05, ** p ≤ 0.01.

    Article Snippet: Drugs and Inhibitors Lapatinib (Lap) (Selleckchem, LubioScience Luzern, Switzerland, S2111) was reconstituted in dimethyl sulfoxide (DMSO) and stock solutions were stored at −80 °C.

    Techniques: Alamar Blue Assay, FACS, Transduction, Construct, MANN-WHITNEY

    Induction of autophagic flux in OE19 upon Lapatinib treatment. ( a ) LC3B flux was assessed comparing control and BafilomycinA (BafA)-treated (200 nM, 2 h) OE19 upon Lapatinib treatment (120 nM, 24 h). LC3 band intensities were quantified using ImageJ software. Total protein was used as a loading control, and Phospho-Her2 for Lapatinib treatment ( n = 3). ( b ) LC3B flux was calculated from data in ( a ) as follows: BafA + -BafA − values for each condition. ( c ) FACS analysis of mCherry-EGFP-LC3B-expressing OE19 cells upon induction or blockade of autophagy with indication of the chosen cut-off value for high respectively low autophagic flux. ( d ) Quantification of the FACS analysis showing % of cells with high autophagic flux ( n = 3). Cells were treated as in ( a ). The error bars represent SD, statistical significance was determined by Mann–Whitney U test: * p ≤ 0.05, ** p ≤ 0.01.

    Journal: International Journal of Molecular Sciences

    Article Title: Her2-Targeted Therapy Induces Autophagy in Esophageal Adenocarcinoma Cells

    doi: 10.3390/ijms19103069

    Figure Lengend Snippet: Induction of autophagic flux in OE19 upon Lapatinib treatment. ( a ) LC3B flux was assessed comparing control and BafilomycinA (BafA)-treated (200 nM, 2 h) OE19 upon Lapatinib treatment (120 nM, 24 h). LC3 band intensities were quantified using ImageJ software. Total protein was used as a loading control, and Phospho-Her2 for Lapatinib treatment ( n = 3). ( b ) LC3B flux was calculated from data in ( a ) as follows: BafA + -BafA − values for each condition. ( c ) FACS analysis of mCherry-EGFP-LC3B-expressing OE19 cells upon induction or blockade of autophagy with indication of the chosen cut-off value for high respectively low autophagic flux. ( d ) Quantification of the FACS analysis showing % of cells with high autophagic flux ( n = 3). Cells were treated as in ( a ). The error bars represent SD, statistical significance was determined by Mann–Whitney U test: * p ≤ 0.05, ** p ≤ 0.01.

    Article Snippet: Drugs and Inhibitors Lapatinib (Lap) (Selleckchem, LubioScience Luzern, Switzerland, S2111) was reconstituted in dimethyl sulfoxide (DMSO) and stock solutions were stored at −80 °C.

    Techniques: Software, FACS, Expressing, MANN-WHITNEY

    Combined Her2 and autophagy inhibitor treatment ( a ) Relative cell viability of OE19 cells treated with Lapatinib (120 nM) and/or autophagy inhibition either VPS34 inhibitor (VPS34-IN1) (5 µM) or chloroquine (CQ) (25 µM) at days 0 and 3 of the alamarBlue ® experiment ( n ≥ 3). ( b ) Cell counts of OE19 P and OE19 LR cells treated as in ( a ) but for 48 h. Values were normalized to DMSO control treated cells ( n = 7) ( c ) Colony numbers after treatment as described in ( b ), reseeded (2500 cells/well) in six-well plates and incubation for 14 days without treatment ( n = 5). The error bars represent SD, statistical significance was determined by Mann–Whitney U test: ns = non significant p > 0.05, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.

    Journal: International Journal of Molecular Sciences

    Article Title: Her2-Targeted Therapy Induces Autophagy in Esophageal Adenocarcinoma Cells

    doi: 10.3390/ijms19103069

    Figure Lengend Snippet: Combined Her2 and autophagy inhibitor treatment ( a ) Relative cell viability of OE19 cells treated with Lapatinib (120 nM) and/or autophagy inhibition either VPS34 inhibitor (VPS34-IN1) (5 µM) or chloroquine (CQ) (25 µM) at days 0 and 3 of the alamarBlue ® experiment ( n ≥ 3). ( b ) Cell counts of OE19 P and OE19 LR cells treated as in ( a ) but for 48 h. Values were normalized to DMSO control treated cells ( n = 7) ( c ) Colony numbers after treatment as described in ( b ), reseeded (2500 cells/well) in six-well plates and incubation for 14 days without treatment ( n = 5). The error bars represent SD, statistical significance was determined by Mann–Whitney U test: ns = non significant p > 0.05, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.

    Article Snippet: Drugs and Inhibitors Lapatinib (Lap) (Selleckchem, LubioScience Luzern, Switzerland, S2111) was reconstituted in dimethyl sulfoxide (DMSO) and stock solutions were stored at −80 °C.

    Techniques: Inhibition, Incubation, MANN-WHITNEY

    A . Western blots of SKBR3 cells transduced with retroviruses encoding either PIK3CA H1047R or E545K mutant alleles or control vector (mCherry). Mutant transduced lines show increased levels of p-AKT and p-S6RP at baseline, and diminished response to lapatinib alone as measured by p-S6RP levels. Note that mutation has little effect on p-4EBP1 levels. B . Growth curves for control, E545K mutant, or H1047R mutant transduced SKBR3 cells treated with lapatinib (red), GSK690693 (green), or a combination of the two (blue). Introduction of mutations results in increased resistance to lapatinib and synergistic interactions between lapatinib and GSK690693 (doses with significant synergy are marked with a red asterisk), indicating a functional role for PIK3CA mutations in determining synergistic response to this combination. C . Analysis of RPPA time course data shows blunted recovery of p-HER3 levels in SKBR3 cells harboring PIK3CA mutations, while the control cell line shows the expected hyper-phosphorylation following lapatinib treatment. D . Western blotting confirms the lack of recovery in non-engineered cells lines with PIK3CA mutations compared to PIK3CA wild type lines.

    Journal: PLoS ONE

    Article Title: Decoupling of the PI3K Pathway via Mutation Necessitates Combinatorial Treatment in HER2+ Breast Cancer

    doi: 10.1371/journal.pone.0133219

    Figure Lengend Snippet: A . Western blots of SKBR3 cells transduced with retroviruses encoding either PIK3CA H1047R or E545K mutant alleles or control vector (mCherry). Mutant transduced lines show increased levels of p-AKT and p-S6RP at baseline, and diminished response to lapatinib alone as measured by p-S6RP levels. Note that mutation has little effect on p-4EBP1 levels. B . Growth curves for control, E545K mutant, or H1047R mutant transduced SKBR3 cells treated with lapatinib (red), GSK690693 (green), or a combination of the two (blue). Introduction of mutations results in increased resistance to lapatinib and synergistic interactions between lapatinib and GSK690693 (doses with significant synergy are marked with a red asterisk), indicating a functional role for PIK3CA mutations in determining synergistic response to this combination. C . Analysis of RPPA time course data shows blunted recovery of p-HER3 levels in SKBR3 cells harboring PIK3CA mutations, while the control cell line shows the expected hyper-phosphorylation following lapatinib treatment. D . Western blotting confirms the lack of recovery in non-engineered cells lines with PIK3CA mutations compared to PIK3CA wild type lines.

    Article Snippet: Additional Lapatinib and GSK690693 for the PIK3CA mutation studies were purchased from Selleck chemicals.

    Techniques: Western Blot, Transduction, Mutagenesis, Plasmid Preparation, Functional Assay

    Examples of A. synergistic (in SUM190PT) and B. antagonistic (in AU565) interactions between lapatinib (red curve) and GSK690693 (green curve) in breast cell lines. Combination treatment is shown in the blue curve.

    Journal: PLoS ONE

    Article Title: Decoupling of the PI3K Pathway via Mutation Necessitates Combinatorial Treatment in HER2+ Breast Cancer

    doi: 10.1371/journal.pone.0133219

    Figure Lengend Snippet: Examples of A. synergistic (in SUM190PT) and B. antagonistic (in AU565) interactions between lapatinib (red curve) and GSK690693 (green curve) in breast cell lines. Combination treatment is shown in the blue curve.

    Article Snippet: Additional Lapatinib and GSK690693 for the PIK3CA mutation studies were purchased from Selleck chemicals.

    Techniques:

    A . A simplified kinetic model for AKT pathway dynamics. Measured protein species (green; yellow circles indicate phosphorylation) are shown along with inhibitors (purple) and edges indicating regulatory interplay. An ERBB independent route to AKT activation is modeled as a node

    Journal: PLoS ONE

    Article Title: Decoupling of the PI3K Pathway via Mutation Necessitates Combinatorial Treatment in HER2+ Breast Cancer

    doi: 10.1371/journal.pone.0133219

    Figure Lengend Snippet: A . A simplified kinetic model for AKT pathway dynamics. Measured protein species (green; yellow circles indicate phosphorylation) are shown along with inhibitors (purple) and edges indicating regulatory interplay. An ERBB independent route to AKT activation is modeled as a node "M" (red) that can activate AKT in PIK3CA mutants only. To allow for the possibility that relevant player(s) were not observed in the RPPA data, the node was formally treated as an unobserved or latent variable. The edge from GSK690693 to p-AKT indicates that GSK690693 abrogates a negative feedback mechanism by which p-AKT regulates its own expression (possibly indirectly). All kinetic parameters were estimated using RPPA time-course data using a Bayesian statistical formulation. Note that links shown are not intended to represent direct influences but rather regulation via intermediate steps that are not explicitly modeled. B . AKT pathway activation (measured as equilibrium level of phospho-S6) as a function of the extent of ERBB (x-axis) and AKT inhibition (y-axis), as predicted by fitted model in A . In the wild type case (upper panel of B ), ERBB inhibition alone is sufficient to abrogate S6 phosphorylation. This is evident by the rapid downregulation of p-S6 with increasing concentrations of lapatinib, such that by the time 80% of ERBB activity is inhibited, there is almost complete inhibition of p-S6. In contrast, PIK3CA mutant lines (lower panel of B) , which have on average higher p-S6 baseline activity than their wild type counterparts, show diminished effects on p-S6 activity with increasing concentrations of lapatinib monotherapy. Indeed, even when ERBB is completely inhibited, there is residual p-S6 activity present. In order to achieve full inhibition of p-S6, combined inhibition of ERBB and AKT is required. For example, to achieve full inhibition of p-S6, the model predicts that inhibition of both ERBB and AKT activity by at least 80% is required in the mutant lines.

    Article Snippet: Additional Lapatinib and GSK690693 for the PIK3CA mutation studies were purchased from Selleck chemicals.

    Techniques: Activation Assay, Expressing, Inhibition, Activity Assay, Mutagenesis

    Heat maps showing average response within subtype (HER2 + /PIK3CA WT ; HER2 + /PIK3CA mut ; HER2 - ) to targeted therapeutics alone and in combination from RPPA data. A . Down-regulation of the PI3K-AKT pathway is evident in all HER2 + cells treated with lapatinib, irrespective of PI3K mutation status. However, the degree of down-regulation is less in cell lines with PI3K mutations (see p-S6 Ser235 for example). B . Treatment with GSK690693 leads to down-regulation of proteins downstream of AKT in all HER2 + cell lines, although the degree of down-regulation is less in lines harboring PI3K mutations. C . Treatment with the combination of lapatinib plus GSK690693 leads to down-regulation of the PI3K-AKT signaling pathway in both PIK3CA mut and PIK3CA WT lines. Note the much greater degree of down-regulation of S235 and S240 p-S6 in PIK3CA mut cell lines treated with the combination relative to the single therapies. D . A summary of the most significant changes observed in the RPPA data for the PIK3CA WT , PIK3CA mut , and non-HER2 amplified cell lines under inhibition with lapatinib, GSK690693, or a combination of the two.

    Journal: PLoS ONE

    Article Title: Decoupling of the PI3K Pathway via Mutation Necessitates Combinatorial Treatment in HER2+ Breast Cancer

    doi: 10.1371/journal.pone.0133219

    Figure Lengend Snippet: Heat maps showing average response within subtype (HER2 + /PIK3CA WT ; HER2 + /PIK3CA mut ; HER2 - ) to targeted therapeutics alone and in combination from RPPA data. A . Down-regulation of the PI3K-AKT pathway is evident in all HER2 + cells treated with lapatinib, irrespective of PI3K mutation status. However, the degree of down-regulation is less in cell lines with PI3K mutations (see p-S6 Ser235 for example). B . Treatment with GSK690693 leads to down-regulation of proteins downstream of AKT in all HER2 + cell lines, although the degree of down-regulation is less in lines harboring PI3K mutations. C . Treatment with the combination of lapatinib plus GSK690693 leads to down-regulation of the PI3K-AKT signaling pathway in both PIK3CA mut and PIK3CA WT lines. Note the much greater degree of down-regulation of S235 and S240 p-S6 in PIK3CA mut cell lines treated with the combination relative to the single therapies. D . A summary of the most significant changes observed in the RPPA data for the PIK3CA WT , PIK3CA mut , and non-HER2 amplified cell lines under inhibition with lapatinib, GSK690693, or a combination of the two.

    Article Snippet: Additional Lapatinib and GSK690693 for the PIK3CA mutation studies were purchased from Selleck chemicals.

    Techniques: Mutagenesis, Amplification, Inhibition

    Effect of AXL and HER2 inhibitors on ESCC cells A. Dose-inhibition curves of CE48T ESCC cells in response to indicated concentration of lapatinib, foretinib, or foretinib plus lapatinib. The respective IC 50 values were 1.891 μM, 0.443 μM and 0.296 μM. B-D. Synergistic effects of foretinib and HER2 inhibitors on the cytotoxicities of ESCC. Cell viability of ESCC cells treated by 1 μM of foretinib, or 1 μM of HER2 inhibitors, or foretinib plus HER2 inhibitor. The HER2 inhibitors included 1 μM of lapatinib B. or 1 μM of afatinib C. or 1 μM of AC480 D. E. Relative cell viabilities of Het-1A and CE48T cells treated with indicated concentration of foretinib. The IC 50 values were 3.836 μM vs. 0.823 μM for Het-1A and CE48T cells respectively. F. Dose-response curves for HER2-sensitive (HER2-S) and HER2-resistant (HER2-R) ESCC cells in response to increased concentrations of lapatinib or foretinib.

    Journal: Oncotarget

    Article Title: The AXL receptor tyrosine kinase is associated with adverse prognosis and distant metastasis in esophageal squamous cell carcinoma

    doi: 10.18632/oncotarget.9231

    Figure Lengend Snippet: Effect of AXL and HER2 inhibitors on ESCC cells A. Dose-inhibition curves of CE48T ESCC cells in response to indicated concentration of lapatinib, foretinib, or foretinib plus lapatinib. The respective IC 50 values were 1.891 μM, 0.443 μM and 0.296 μM. B-D. Synergistic effects of foretinib and HER2 inhibitors on the cytotoxicities of ESCC. Cell viability of ESCC cells treated by 1 μM of foretinib, or 1 μM of HER2 inhibitors, or foretinib plus HER2 inhibitor. The HER2 inhibitors included 1 μM of lapatinib B. or 1 μM of afatinib C. or 1 μM of AC480 D. E. Relative cell viabilities of Het-1A and CE48T cells treated with indicated concentration of foretinib. The IC 50 values were 3.836 μM vs. 0.823 μM for Het-1A and CE48T cells respectively. F. Dose-response curves for HER2-sensitive (HER2-S) and HER2-resistant (HER2-R) ESCC cells in response to increased concentrations of lapatinib or foretinib.

    Article Snippet: Isolation of HER2-resistant ESCC cells HER2-resistant ESCC sub-cells were selected by repeated treatment with the HER2 inhibitor lapatinib (Tykerb, synthesized by Selleckchem, S2111) [ ] of different concentrations (5 to 20 μM/L).

    Techniques: Inhibition, Concentration Assay

    A. Protein expression profiles of CE48T, HER2-resistant CE48T cells (CE48T-HER2-R) and Het-1A cells. α-Tubulin served as a loading control for phospho-ERK (pERK) and ERK whereas β-Actin was used as a loading control for AXL, HER2, phospho-AKT (pAKT), and AKT. B. Phospho-HER2 (pHER2), HER2, pERK, and ERK expression in ESCC cells treated with indicated amounts of lapatinib for 24 hours. β-Actin served as a loading control. C. Phospho-AXL (pAXL), AXL, pERK, ERK, pAKT, and AKT expression in ESCC cells treated with the indicated amounts of foretinib for 24 hours. α-Tubulin served as a loading control for pERK and ERK; β-Actin served as a loading control for pAXL, AXL, pAKT, and AKT.

    Journal: Oncotarget

    Article Title: The AXL receptor tyrosine kinase is associated with adverse prognosis and distant metastasis in esophageal squamous cell carcinoma

    doi: 10.18632/oncotarget.9231

    Figure Lengend Snippet: A. Protein expression profiles of CE48T, HER2-resistant CE48T cells (CE48T-HER2-R) and Het-1A cells. α-Tubulin served as a loading control for phospho-ERK (pERK) and ERK whereas β-Actin was used as a loading control for AXL, HER2, phospho-AKT (pAKT), and AKT. B. Phospho-HER2 (pHER2), HER2, pERK, and ERK expression in ESCC cells treated with indicated amounts of lapatinib for 24 hours. β-Actin served as a loading control. C. Phospho-AXL (pAXL), AXL, pERK, ERK, pAKT, and AKT expression in ESCC cells treated with the indicated amounts of foretinib for 24 hours. α-Tubulin served as a loading control for pERK and ERK; β-Actin served as a loading control for pAXL, AXL, pAKT, and AKT.

    Article Snippet: Isolation of HER2-resistant ESCC cells HER2-resistant ESCC sub-cells were selected by repeated treatment with the HER2 inhibitor lapatinib (Tykerb, synthesized by Selleckchem, S2111) [ ] of different concentrations (5 to 20 μM/L).

    Techniques: Expressing

    Schematic model of the JWA/c-Cbl/HER2 pathway's role in lapatinib resistance Left: low expression of JWA contributes to HER2 stabilization and lapatinib sensitivity. Right: JWA overexpression promotes the E3 ubiquitin ligase c-Cbl, leading to an increase in Her2 polyubiquitination. This ultimately results in a decrease in Her2 protein levels and confers lapatinib resistance.

    Journal: Oncotarget

    Article Title: JWA down-regulates HER2 expression via c-Cbl and induces lapatinib resistance in human gastric cancer cells

    doi: 10.18632/oncotarget.12374

    Figure Lengend Snippet: Schematic model of the JWA/c-Cbl/HER2 pathway's role in lapatinib resistance Left: low expression of JWA contributes to HER2 stabilization and lapatinib sensitivity. Right: JWA overexpression promotes the E3 ubiquitin ligase c-Cbl, leading to an increase in Her2 polyubiquitination. This ultimately results in a decrease in Her2 protein levels and confers lapatinib resistance.

    Article Snippet: Drug preparations and reagents Lapatinib (Selleck Chemicals, USA) was dissolved in 100% dimethyl sulfoxide (DMSO; Sigma-Aldrich, USA) and then diluted with culture medium to the desired concentration.

    Techniques: Expressing, Over Expression

    HER2 level contributes to lapatinib sensitivity ( A ) The cell viability was measured by CCK8 assay. SGC-7901 cells were exposed to different concentrations of lapatinib for 24 h after transfection with pcDNA3.0 or HER2-WT plasmid for 48 h. ( B ) NCI-N87 cells transfected with or without HER2 siRNA were treated with varying concentrations of lapatinib for 24 hours. The cell survival rates are expressed as means ± SD from at least three independent experiments. * P

    Journal: Oncotarget

    Article Title: JWA down-regulates HER2 expression via c-Cbl and induces lapatinib resistance in human gastric cancer cells

    doi: 10.18632/oncotarget.12374

    Figure Lengend Snippet: HER2 level contributes to lapatinib sensitivity ( A ) The cell viability was measured by CCK8 assay. SGC-7901 cells were exposed to different concentrations of lapatinib for 24 h after transfection with pcDNA3.0 or HER2-WT plasmid for 48 h. ( B ) NCI-N87 cells transfected with or without HER2 siRNA were treated with varying concentrations of lapatinib for 24 hours. The cell survival rates are expressed as means ± SD from at least three independent experiments. * P

    Article Snippet: Drug preparations and reagents Lapatinib (Selleck Chemicals, USA) was dissolved in 100% dimethyl sulfoxide (DMSO; Sigma-Aldrich, USA) and then diluted with culture medium to the desired concentration.

    Techniques: CCK-8 Assay, Transfection, Plasmid Preparation

    JWA decreases the sensitivity of GC cells to lapatinib ( A ) Expressions of HER2 and JWA were examined in whole-cell lysates by Western blotting. ( B and C ) SGC-7901 cells with or without JWA knockdown (B) and NCI-N87 cells with or without JWA overexpression (C) were treated with the indicated doses of lapatinib for 24 h. Cell survival was determined using the CCK8 assay. The cell survival rates are presented as means ± SD from three independent experiments. ( D ) SGC-7901 cells were transfected with si-JWA or its vector for 48 h, followed by incubation with 30 μM lapatinib for 24 h, and then analyzed by flow cytometry. ( E ) NCI-N87 cells were transfected with Flag-JWA or its vector for 48 h, followed by incubation with 1 μM lapatinib for 24 h, and then analyzed by flow cytometry. ( F and G ) Quantification of apoptosis in D and E. Columns indicate average of triplicates and bars indicate SD. * P

    Journal: Oncotarget

    Article Title: JWA down-regulates HER2 expression via c-Cbl and induces lapatinib resistance in human gastric cancer cells

    doi: 10.18632/oncotarget.12374

    Figure Lengend Snippet: JWA decreases the sensitivity of GC cells to lapatinib ( A ) Expressions of HER2 and JWA were examined in whole-cell lysates by Western blotting. ( B and C ) SGC-7901 cells with or without JWA knockdown (B) and NCI-N87 cells with or without JWA overexpression (C) were treated with the indicated doses of lapatinib for 24 h. Cell survival was determined using the CCK8 assay. The cell survival rates are presented as means ± SD from three independent experiments. ( D ) SGC-7901 cells were transfected with si-JWA or its vector for 48 h, followed by incubation with 30 μM lapatinib for 24 h, and then analyzed by flow cytometry. ( E ) NCI-N87 cells were transfected with Flag-JWA or its vector for 48 h, followed by incubation with 1 μM lapatinib for 24 h, and then analyzed by flow cytometry. ( F and G ) Quantification of apoptosis in D and E. Columns indicate average of triplicates and bars indicate SD. * P

    Article Snippet: Drug preparations and reagents Lapatinib (Selleck Chemicals, USA) was dissolved in 100% dimethyl sulfoxide (DMSO; Sigma-Aldrich, USA) and then diluted with culture medium to the desired concentration.

    Techniques: Western Blot, Over Expression, CCK-8 Assay, Transfection, Plasmid Preparation, Incubation, Flow Cytometry, Cytometry

    JWA mediates lapatinib resistance by negatively regulating HER2 ( A ) SGC-7901 cells were transfected with JWA siRNA or 48 h, followed by exposure to 30 μM lapatinib for 24 h. The apoptotic rate was determined by the TUNEL assay (×1000). ( B ) Quantification of TUNEL-positive SGC-7901 cells transfected with JWA siRNA. ( C ) SGC-7901 cells were treated with lapatinib as in (A) and whole-cell lysates were collected for detection of target proteins by Western blotting. ( D ) NCI-N87 cells were transfected with Flag-JWA and then treated with 1μM lapatinib for 24 h. The apoptotic rate was determined by the TUNEL assay (× 1000). ( E ) Quantification of TUNEL-positive NCI-N87 cells transfected with Flag-JWA. Columns indicate average of triplicates and bars indicate SD. * P

    Journal: Oncotarget

    Article Title: JWA down-regulates HER2 expression via c-Cbl and induces lapatinib resistance in human gastric cancer cells

    doi: 10.18632/oncotarget.12374

    Figure Lengend Snippet: JWA mediates lapatinib resistance by negatively regulating HER2 ( A ) SGC-7901 cells were transfected with JWA siRNA or 48 h, followed by exposure to 30 μM lapatinib for 24 h. The apoptotic rate was determined by the TUNEL assay (×1000). ( B ) Quantification of TUNEL-positive SGC-7901 cells transfected with JWA siRNA. ( C ) SGC-7901 cells were treated with lapatinib as in (A) and whole-cell lysates were collected for detection of target proteins by Western blotting. ( D ) NCI-N87 cells were transfected with Flag-JWA and then treated with 1μM lapatinib for 24 h. The apoptotic rate was determined by the TUNEL assay (× 1000). ( E ) Quantification of TUNEL-positive NCI-N87 cells transfected with Flag-JWA. Columns indicate average of triplicates and bars indicate SD. * P

    Article Snippet: Drug preparations and reagents Lapatinib (Selleck Chemicals, USA) was dissolved in 100% dimethyl sulfoxide (DMSO; Sigma-Aldrich, USA) and then diluted with culture medium to the desired concentration.

    Techniques: Transfection, TUNEL Assay, Western Blot

    JWA negatively regulates HER2 expression via c-Cbl in GC cells ( A ) Western blotting was used to determine the expressions of HER2, c-Cbl and JWA in SGC-7901 and NCI-N87 cells. ( B ) SGC-7901 and NCI-N87 were transfected with Flag-JWA or JWA siRNA for 48 h, target proteins were determined with Western blot. ( C ) SGC-7901 cells were transfected with c-Cbl siRNA for 48 h, and western blotting showed the expression of target proteins. ( D ) NCI-N87 were transfected with Flag-Ctrl (top), Flag-JWA (middle), or cotransfected with Flag-JWA and c-Cbl siRNA (bottom) for 48 h. Immunofluorescence imaging of HER2 (green), the lysosome marker Lamp2 (red), nucleus labeled as DAPI (blue), the co-localization of the three signals (merge). ( E ) NCI-N87 cells were cotransfected with Flag-JWA and c-Cbl siRNA for 48 h, incubated with or without lapatinib (1 μM) for 24 h. Western blotting was used to determine the expression of target proteins.

    Journal: Oncotarget

    Article Title: JWA down-regulates HER2 expression via c-Cbl and induces lapatinib resistance in human gastric cancer cells

    doi: 10.18632/oncotarget.12374

    Figure Lengend Snippet: JWA negatively regulates HER2 expression via c-Cbl in GC cells ( A ) Western blotting was used to determine the expressions of HER2, c-Cbl and JWA in SGC-7901 and NCI-N87 cells. ( B ) SGC-7901 and NCI-N87 were transfected with Flag-JWA or JWA siRNA for 48 h, target proteins were determined with Western blot. ( C ) SGC-7901 cells were transfected with c-Cbl siRNA for 48 h, and western blotting showed the expression of target proteins. ( D ) NCI-N87 were transfected with Flag-Ctrl (top), Flag-JWA (middle), or cotransfected with Flag-JWA and c-Cbl siRNA (bottom) for 48 h. Immunofluorescence imaging of HER2 (green), the lysosome marker Lamp2 (red), nucleus labeled as DAPI (blue), the co-localization of the three signals (merge). ( E ) NCI-N87 cells were cotransfected with Flag-JWA and c-Cbl siRNA for 48 h, incubated with or without lapatinib (1 μM) for 24 h. Western blotting was used to determine the expression of target proteins.

    Article Snippet: Drug preparations and reagents Lapatinib (Selleck Chemicals, USA) was dissolved in 100% dimethyl sulfoxide (DMSO; Sigma-Aldrich, USA) and then diluted with culture medium to the desired concentration.

    Techniques: Expressing, Western Blot, Transfection, Immunofluorescence, Imaging, Marker, Labeling, Incubation

    Lapatinib is effective to intrinsic cisplatin-resistant GC cells ( A ) Cell viability was determined by CCK-8 cell-proliferation assay. The gastric cancer (GC) cell lines (BGC-823, SGC-7901, HGC-27, NCI-N87) were exposed to 0.17 μM cisplatin for 24, 48 and 72 h. The percentage of viable cells was shown relative to untreated controls. ( B ) The cell viabilities of four GC cell lines were measured using CCK-8 assay after 48 h exposure to 0, 0.1, 1 and 10 μM of lapatinib. ( C ) Cell death was determined by TUNEL assay (× 1000). Top: The four GC cell lines were treated with 0, 0.17, 1.7 μM cisplatin for 24 h. Bottom: The four GC cell lines were treated with 0, 1, 10 μM lapatinib for 24 h. ( D ) Quantify TUNEL-positive cells of GC cells treated with cisplatin or lapatinib. ( E ) Western blot showing EGFR, HER2 and HER3 protein expressions in the four GC cell lines. The data presented are means ± SD from three independent experiments. * P

    Journal: Oncotarget

    Article Title: JWA down-regulates HER2 expression via c-Cbl and induces lapatinib resistance in human gastric cancer cells

    doi: 10.18632/oncotarget.12374

    Figure Lengend Snippet: Lapatinib is effective to intrinsic cisplatin-resistant GC cells ( A ) Cell viability was determined by CCK-8 cell-proliferation assay. The gastric cancer (GC) cell lines (BGC-823, SGC-7901, HGC-27, NCI-N87) were exposed to 0.17 μM cisplatin for 24, 48 and 72 h. The percentage of viable cells was shown relative to untreated controls. ( B ) The cell viabilities of four GC cell lines were measured using CCK-8 assay after 48 h exposure to 0, 0.1, 1 and 10 μM of lapatinib. ( C ) Cell death was determined by TUNEL assay (× 1000). Top: The four GC cell lines were treated with 0, 0.17, 1.7 μM cisplatin for 24 h. Bottom: The four GC cell lines were treated with 0, 1, 10 μM lapatinib for 24 h. ( D ) Quantify TUNEL-positive cells of GC cells treated with cisplatin or lapatinib. ( E ) Western blot showing EGFR, HER2 and HER3 protein expressions in the four GC cell lines. The data presented are means ± SD from three independent experiments. * P

    Article Snippet: Drug preparations and reagents Lapatinib (Selleck Chemicals, USA) was dissolved in 100% dimethyl sulfoxide (DMSO; Sigma-Aldrich, USA) and then diluted with culture medium to the desired concentration.

    Techniques: CCK-8 Assay, Proliferation Assay, TUNEL Assay, Western Blot

    Inhibition of EGFR signaling in bladder cancer cells by lapatinib in a co-culture system abrogates co-culture induced cancer cell malignancy and proliferation. (A) Western blot analysis indicates that downstream EGFR signaling was inhibited by co-culture treatment with lapatinib. (B and C) Transwell migration assays indicated that co-culture induced malignancy of T24/253J is attenuated in the presence of lapatinib (magnification, ×200). (D and E) Colony formation assay indicated that the co-culture induced enhanced proliferation of T24/253J is ameliorated in the absence of EGFR signaling (magnification, ×200). (F and G) MTT assay determination of bladder cancer T24/253J cell proliferation following co-culture treatment with lapatinib. (H) Western blot analysis indicates that in co-culture system the proteins MMP2, MMP9, ZEB-1, survivin and N-cadherin were downregulated by inhibited EGFR signaling. *** P

    Journal: International Journal of Oncology

    Article Title: Bladder cancer cells interact with vascular endothelial cells triggering EGFR signals to promote tumor progression

    doi: 10.3892/ijo.2019.4729

    Figure Lengend Snippet: Inhibition of EGFR signaling in bladder cancer cells by lapatinib in a co-culture system abrogates co-culture induced cancer cell malignancy and proliferation. (A) Western blot analysis indicates that downstream EGFR signaling was inhibited by co-culture treatment with lapatinib. (B and C) Transwell migration assays indicated that co-culture induced malignancy of T24/253J is attenuated in the presence of lapatinib (magnification, ×200). (D and E) Colony formation assay indicated that the co-culture induced enhanced proliferation of T24/253J is ameliorated in the absence of EGFR signaling (magnification, ×200). (F and G) MTT assay determination of bladder cancer T24/253J cell proliferation following co-culture treatment with lapatinib. (H) Western blot analysis indicates that in co-culture system the proteins MMP2, MMP9, ZEB-1, survivin and N-cadherin were downregulated by inhibited EGFR signaling. *** P

    Article Snippet: In order to mimic the interaction between a cancer cell and EC in the tumor microenvironment, 0.4-µ m pore diameter chambers of a 6-well plate (EMD Millipore, Billerica, MA, USA) were used; 50,000 T24/253J cells were added to the lower chambers of the 6-well plate, and 50,000 HUVECs were added to the upper chambers, and cultured in DMEM (Invitrogen; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Invitrogen; Thermo Fisher Scientific, Inc.) in an atmosphere with 5% CO2 at 37°C for 72 h. The EGFR inhibitor lapatinib, the VEGFR inhibitor ZM 323881 HCl and the CXCR2 inhibitor SB225002 were obtained from Selleck Chemicals (Houston, TX, USA); the NF-κB inhibitor PDTC was obtained from Sigma-Aldrich; Merck KGaA (Darmstadt, Germany), dissolved in dimethyl sulfoxide (DMSO) and stored at −20°C.

    Techniques: Inhibition, Co-Culture Assay, Western Blot, Migration, Colony Assay, MTT Assay

    Diagram proposing a model for the interaction between bladder cancer cells and endothelial cells. In the co-culture system, both bladder cancer cells and endothelial cells secrete the VEGFR2 ligands VEGF-A and VEGF-C, which induce VEGFR2 signaling and downstream NF-κB signaling, promoting EGFR ligand expression. These events may be inhibited by a VEGFR2 inhibitor, ZM and an NF-κB inhibitor (PDTC). EGFR signaling in bladder cancer cells was triggered by EGFR ligands secreted by endothelial cells, which induces phosphorylation of AKT and NF-κB. These events enhance bladder cancer migration, invasion, and proliferation. Furthermore, activated EGFR signaling in bladder cancer cells could enhance endothelial cell recruitment through the upregulation of CXCL1, CXCL5 and CXCL8. These events could be inhibited by an EGFR inhibitor, lap, PDTC and a CXCR2 inhibitor, SB. VEGF, vascular endothelial growth factor; R, receptor; NF, nuclear factor; EGFR, epidermal growth factor receptor; ZM, ZM 323881 HCL; AKT, protein kinase B; lap, lapatinib; SB, SB225002.

    Journal: International Journal of Oncology

    Article Title: Bladder cancer cells interact with vascular endothelial cells triggering EGFR signals to promote tumor progression

    doi: 10.3892/ijo.2019.4729

    Figure Lengend Snippet: Diagram proposing a model for the interaction between bladder cancer cells and endothelial cells. In the co-culture system, both bladder cancer cells and endothelial cells secrete the VEGFR2 ligands VEGF-A and VEGF-C, which induce VEGFR2 signaling and downstream NF-κB signaling, promoting EGFR ligand expression. These events may be inhibited by a VEGFR2 inhibitor, ZM and an NF-κB inhibitor (PDTC). EGFR signaling in bladder cancer cells was triggered by EGFR ligands secreted by endothelial cells, which induces phosphorylation of AKT and NF-κB. These events enhance bladder cancer migration, invasion, and proliferation. Furthermore, activated EGFR signaling in bladder cancer cells could enhance endothelial cell recruitment through the upregulation of CXCL1, CXCL5 and CXCL8. These events could be inhibited by an EGFR inhibitor, lap, PDTC and a CXCR2 inhibitor, SB. VEGF, vascular endothelial growth factor; R, receptor; NF, nuclear factor; EGFR, epidermal growth factor receptor; ZM, ZM 323881 HCL; AKT, protein kinase B; lap, lapatinib; SB, SB225002.

    Article Snippet: In order to mimic the interaction between a cancer cell and EC in the tumor microenvironment, 0.4-µ m pore diameter chambers of a 6-well plate (EMD Millipore, Billerica, MA, USA) were used; 50,000 T24/253J cells were added to the lower chambers of the 6-well plate, and 50,000 HUVECs were added to the upper chambers, and cultured in DMEM (Invitrogen; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Invitrogen; Thermo Fisher Scientific, Inc.) in an atmosphere with 5% CO2 at 37°C for 72 h. The EGFR inhibitor lapatinib, the VEGFR inhibitor ZM 323881 HCl and the CXCR2 inhibitor SB225002 were obtained from Selleck Chemicals (Houston, TX, USA); the NF-κB inhibitor PDTC was obtained from Sigma-Aldrich; Merck KGaA (Darmstadt, Germany), dissolved in dimethyl sulfoxide (DMSO) and stored at −20°C.

    Techniques: Co-Culture Assay, Expressing, Migration

    The inhibitory effect of lapatinib on the proliferation of 22 cancer cell lines and the rescue effect of fibroblast culture supernatant against lapatinib-induced inhibition. Lapanitib (1 μM) inhibits cell proliferation by more than 50% in the majority of the ESCC cell lines. Addition of fibroblast supernatant rescues growth inhibition by lapatinib and recovers cell proliferation by 20% or more in 10 cell lines.

    Journal: BMC Cancer

    Article Title: The role of HGF/MET and FGF/FGFR in fibroblast-derived growth stimulation and lapatinib-resistance of esophageal squamous cell carcinoma

    doi: 10.1186/s12885-015-1065-8

    Figure Lengend Snippet: The inhibitory effect of lapatinib on the proliferation of 22 cancer cell lines and the rescue effect of fibroblast culture supernatant against lapatinib-induced inhibition. Lapanitib (1 μM) inhibits cell proliferation by more than 50% in the majority of the ESCC cell lines. Addition of fibroblast supernatant rescues growth inhibition by lapatinib and recovers cell proliferation by 20% or more in 10 cell lines.

    Article Snippet: Lapatinib was obtained from Bio Vision (Milpitas, USA).

    Techniques: Inhibition

    Rescue from lapatinib inhibition by FB-SN and abrogation by FGFR inhibitor. The inhibitory effect of lapatinib on the proliferation of representative cell lines is rescued by the addition of fibroblast culture supernatant. Further addition of FGFR inhibitor (PD-173074) abrogates the rescue effect of fibroblast supernatant. The Y axis indicates the absorbance.

    Journal: BMC Cancer

    Article Title: The role of HGF/MET and FGF/FGFR in fibroblast-derived growth stimulation and lapatinib-resistance of esophageal squamous cell carcinoma

    doi: 10.1186/s12885-015-1065-8

    Figure Lengend Snippet: Rescue from lapatinib inhibition by FB-SN and abrogation by FGFR inhibitor. The inhibitory effect of lapatinib on the proliferation of representative cell lines is rescued by the addition of fibroblast culture supernatant. Further addition of FGFR inhibitor (PD-173074) abrogates the rescue effect of fibroblast supernatant. The Y axis indicates the absorbance.

    Article Snippet: Lapatinib was obtained from Bio Vision (Milpitas, USA).

    Techniques: Inhibition

    BT474 lapatinib resistant cells with prolonged treatment reactivate HER receptor activity . (A) Growth curves of UACC-812 and BT474 late stage lapatinib resistant cells (LLR) treated with different targeted therapies for nine days: trastuzumab (T) (10 μg/ml), lapatinib (L) (1 μM), trastuzumab plus lapatinib, or endocrine therapy, fulvestrant (F) (10 -7 M). Significance between groups was determined by multiple comparisons using the Sidak method (* P = 0.0008, BT474 LLR + L versus LLR + L + F, * P = 0.0044, BT474 LLR + L versus LLR + L + T; * P

    Journal: Breast Cancer Research : BCR

    Article Title: Different mechanisms for resistance to trastuzumab versus lapatinib in HER2- positive breast cancers -- role of estrogen receptor and HER2 reactivation

    doi: 10.1186/bcr3067

    Figure Lengend Snippet: BT474 lapatinib resistant cells with prolonged treatment reactivate HER receptor activity . (A) Growth curves of UACC-812 and BT474 late stage lapatinib resistant cells (LLR) treated with different targeted therapies for nine days: trastuzumab (T) (10 μg/ml), lapatinib (L) (1 μM), trastuzumab plus lapatinib, or endocrine therapy, fulvestrant (F) (10 -7 M). Significance between groups was determined by multiple comparisons using the Sidak method (* P = 0.0008, BT474 LLR + L versus LLR + L + F, * P = 0.0044, BT474 LLR + L versus LLR + L + T; * P

    Article Snippet: Lapatinib (Tykerb) was obtained from GlaxoSmithKline (US headquarters in Research Triangle Park, NC, USA) and prepared with dimethyl sulfoxide (DMSO).

    Techniques: Activity Assay

    Inhibition of HER2 restores lapatinib sensitivity in BT474 late stage lapatinib resistant cells . (A) BT474 parental and resistant cells were treated with pooled EGFR, HER2, HER3, ER siRNA, or non-targeting control siRNA, for 72 hours. Proliferation was measured using the Click-iT EdU (5-ethynyl-2'- deoxyuridine) Microplate Assay. Apoptosis was measured by detecting Annexin V expression. Signals were visualized and quantitated by the Celigo cytometer (Cyntellect, San Diego, CA, USA). (B) Down-regulation of EGFR, HER2, HER3, and ER in BT474 derivatives after siRNA treatment was detected by Western blot. Whole-cell extracts were analyzed with the indicated antibodies, including downstream signaling. (C) Growth fold change of double dosage (2 μM) lapatinib on BT474 early and late stage-lapatinib resistant cells for six-day treatment. Cell numbers were assessed using methylene blue and quantified by absorbance at 655 nm and normalized against Day 0. Significance between groups was determined by multiple comparisons using the Sidak method (* P

    Journal: Breast Cancer Research : BCR

    Article Title: Different mechanisms for resistance to trastuzumab versus lapatinib in HER2- positive breast cancers -- role of estrogen receptor and HER2 reactivation

    doi: 10.1186/bcr3067

    Figure Lengend Snippet: Inhibition of HER2 restores lapatinib sensitivity in BT474 late stage lapatinib resistant cells . (A) BT474 parental and resistant cells were treated with pooled EGFR, HER2, HER3, ER siRNA, or non-targeting control siRNA, for 72 hours. Proliferation was measured using the Click-iT EdU (5-ethynyl-2'- deoxyuridine) Microplate Assay. Apoptosis was measured by detecting Annexin V expression. Signals were visualized and quantitated by the Celigo cytometer (Cyntellect, San Diego, CA, USA). (B) Down-regulation of EGFR, HER2, HER3, and ER in BT474 derivatives after siRNA treatment was detected by Western blot. Whole-cell extracts were analyzed with the indicated antibodies, including downstream signaling. (C) Growth fold change of double dosage (2 μM) lapatinib on BT474 early and late stage-lapatinib resistant cells for six-day treatment. Cell numbers were assessed using methylene blue and quantified by absorbance at 655 nm and normalized against Day 0. Significance between groups was determined by multiple comparisons using the Sidak method (* P

    Article Snippet: Lapatinib (Tykerb) was obtained from GlaxoSmithKline (US headquarters in Research Triangle Park, NC, USA) and prepared with dimethyl sulfoxide (DMSO).

    Techniques: Inhibition, Expressing, Cytometry, Western Blot

    BT474 late stage lapatinib-resistant cells overexpress HER2 and HER ligands . (A) mRNA expression levels of HER receptors and ligands in BT474 parental and distinct resistant derivatives by qRT-PCR. Data were normalized to parental cells. (B) EGFR, HER2, and HER3 protein levels in BT474 parental, early, and late stage lapatinib-resistant cells. Protein level was quantified with Odyssey software (LI-COR Biosciences, Inc., Lincoln, NE). Each expression level was acquired from three independent samples for each derivative. Significance between groups was determined by multiple comparisons using the Sidak method (* P

    Journal: Breast Cancer Research : BCR

    Article Title: Different mechanisms for resistance to trastuzumab versus lapatinib in HER2- positive breast cancers -- role of estrogen receptor and HER2 reactivation

    doi: 10.1186/bcr3067

    Figure Lengend Snippet: BT474 late stage lapatinib-resistant cells overexpress HER2 and HER ligands . (A) mRNA expression levels of HER receptors and ligands in BT474 parental and distinct resistant derivatives by qRT-PCR. Data were normalized to parental cells. (B) EGFR, HER2, and HER3 protein levels in BT474 parental, early, and late stage lapatinib-resistant cells. Protein level was quantified with Odyssey software (LI-COR Biosciences, Inc., Lincoln, NE). Each expression level was acquired from three independent samples for each derivative. Significance between groups was determined by multiple comparisons using the Sidak method (* P

    Article Snippet: Lapatinib (Tykerb) was obtained from GlaxoSmithKline (US headquarters in Research Triangle Park, NC, USA) and prepared with dimethyl sulfoxide (DMSO).

    Techniques: Expressing, Quantitative RT-PCR, Software

    Resistant cells show greater proliferation and exhibit changes in ER and PR expression . (A) Cell proliferation assay of UACC-812 and BT474 parental and resistant (R) cells. Cells were treated with trastuzumab (T, 10 μg/ml), lapatinib (L, 1 μM), or trastuzumab plus lapatinib (L + T). After six days, viable cells were visualized by methylene blue staining and photographed. (B) Fold changes in cell growth of UACC-812 and BT474 parental and resistant cells with or without the respective anti-HER2 therapies, following six days of treatment. Cell numbers were quantified by absorbance at 655 nm and normalized against Day 0. Significance between groups was determined by multiple comparisons using the Sidak method (* P

    Journal: Breast Cancer Research : BCR

    Article Title: Different mechanisms for resistance to trastuzumab versus lapatinib in HER2- positive breast cancers -- role of estrogen receptor and HER2 reactivation

    doi: 10.1186/bcr3067

    Figure Lengend Snippet: Resistant cells show greater proliferation and exhibit changes in ER and PR expression . (A) Cell proliferation assay of UACC-812 and BT474 parental and resistant (R) cells. Cells were treated with trastuzumab (T, 10 μg/ml), lapatinib (L, 1 μM), or trastuzumab plus lapatinib (L + T). After six days, viable cells were visualized by methylene blue staining and photographed. (B) Fold changes in cell growth of UACC-812 and BT474 parental and resistant cells with or without the respective anti-HER2 therapies, following six days of treatment. Cell numbers were quantified by absorbance at 655 nm and normalized against Day 0. Significance between groups was determined by multiple comparisons using the Sidak method (* P

    Article Snippet: Lapatinib (Tykerb) was obtained from GlaxoSmithKline (US headquarters in Research Triangle Park, NC, USA) and prepared with dimethyl sulfoxide (DMSO).

    Techniques: Expressing, Proliferation Assay, Staining

    Trastuzumab resistant cells maintain HER signaling. Lapatinib and combination resistant cells express up-regulated ER activity . (A) qRT-PCR expression levels of ER and PR mRNA in UACC-812 and BT474 parental and distinct resistant clones. Data were normalized to parental cells. (B) UACC-812 and BT474 parental cells were treated with trastuzumab (10 μg/ml), lapatinib (1 μM), or trastuzumab plus lapatinib for five hours and harvested. Whole-cell extracts of these treatment groups and resistant derivatives were analyzed by Western blot with the indicated antibodies.

    Journal: Breast Cancer Research : BCR

    Article Title: Different mechanisms for resistance to trastuzumab versus lapatinib in HER2- positive breast cancers -- role of estrogen receptor and HER2 reactivation

    doi: 10.1186/bcr3067

    Figure Lengend Snippet: Trastuzumab resistant cells maintain HER signaling. Lapatinib and combination resistant cells express up-regulated ER activity . (A) qRT-PCR expression levels of ER and PR mRNA in UACC-812 and BT474 parental and distinct resistant clones. Data were normalized to parental cells. (B) UACC-812 and BT474 parental cells were treated with trastuzumab (10 μg/ml), lapatinib (1 μM), or trastuzumab plus lapatinib for five hours and harvested. Whole-cell extracts of these treatment groups and resistant derivatives were analyzed by Western blot with the indicated antibodies.

    Article Snippet: Lapatinib (Tykerb) was obtained from GlaxoSmithKline (US headquarters in Research Triangle Park, NC, USA) and prepared with dimethyl sulfoxide (DMSO).

    Techniques: Activity Assay, Quantitative RT-PCR, Expressing, Clone Assay, Western Blot

    Trastuzumab resistant cells remain sensitive to lapatinib. Fulvestrant inhibits lapatinib and combination resistant cell growth . (A) Growth curves of UACC-812 and BT474 parental and resistant cells treated with different target therapies/regimens for nine days: trastuzumab (T) (10 μg/ml), lapatinib (L) (1 μM), trastuzumab plus lapatinib (L + T), or endocrine therapy, fulvestrant (F) (10 -7 M); media of parental cells (C). Cell numbers were quantified by absorbance at 655 nm after staining with methylene blue. Conditions were repeated in quadruplicate. Significance between groups was determined by multiple comparisons using the Sidak method (* P

    Journal: Breast Cancer Research : BCR

    Article Title: Different mechanisms for resistance to trastuzumab versus lapatinib in HER2- positive breast cancers -- role of estrogen receptor and HER2 reactivation

    doi: 10.1186/bcr3067

    Figure Lengend Snippet: Trastuzumab resistant cells remain sensitive to lapatinib. Fulvestrant inhibits lapatinib and combination resistant cell growth . (A) Growth curves of UACC-812 and BT474 parental and resistant cells treated with different target therapies/regimens for nine days: trastuzumab (T) (10 μg/ml), lapatinib (L) (1 μM), trastuzumab plus lapatinib (L + T), or endocrine therapy, fulvestrant (F) (10 -7 M); media of parental cells (C). Cell numbers were quantified by absorbance at 655 nm after staining with methylene blue. Conditions were repeated in quadruplicate. Significance between groups was determined by multiple comparisons using the Sidak method (* P

    Article Snippet: Lapatinib (Tykerb) was obtained from GlaxoSmithKline (US headquarters in Research Triangle Park, NC, USA) and prepared with dimethyl sulfoxide (DMSO).

    Techniques: Staining

    Growth of UACC-812 xenografts treated with various anti-HER2 treatments, with or without estrogen deprivation . (A) Treatment in the presence of estrogen supplementation, representing no endocrine therapy. Treatments included: Estrogen alone (E2) or with lapatinib (E2 + L), trastuzumab (E2 + T), or their combination (E2 + L + T). (B) Treatments in the presence of endocrine therapy in the form of estrogen deprivation. Treatments included: Estrogen (E2), estrogen deprivation (ED) alone, or along with lapatinib (ED + L), trastuzumab (ED + T), or their combination (ED + L + T). Results are presented as the mean tumor volume; error bars represent the standard error.

    Journal: Breast Cancer Research : BCR

    Article Title: Different mechanisms for resistance to trastuzumab versus lapatinib in HER2- positive breast cancers -- role of estrogen receptor and HER2 reactivation

    doi: 10.1186/bcr3067

    Figure Lengend Snippet: Growth of UACC-812 xenografts treated with various anti-HER2 treatments, with or without estrogen deprivation . (A) Treatment in the presence of estrogen supplementation, representing no endocrine therapy. Treatments included: Estrogen alone (E2) or with lapatinib (E2 + L), trastuzumab (E2 + T), or their combination (E2 + L + T). (B) Treatments in the presence of endocrine therapy in the form of estrogen deprivation. Treatments included: Estrogen (E2), estrogen deprivation (ED) alone, or along with lapatinib (ED + L), trastuzumab (ED + T), or their combination (ED + L + T). Results are presented as the mean tumor volume; error bars represent the standard error.

    Article Snippet: Lapatinib (Tykerb) was obtained from GlaxoSmithKline (US headquarters in Research Triangle Park, NC, USA) and prepared with dimethyl sulfoxide (DMSO).

    Techniques:

    HER2-overexpressing cell lines exhibit distinct responses when treated with potent anti-HER2 therapy . (A) A panel of HER2-overexpressing breast cancer cell lines was treated with lapatinib (1 μM) plus trastuzumab (10 μg/ml) for 48 h and whole-cell extracts were analyzed by immunoblotting with indicated antibodies. (B) Combination therapy (trastuzumab plus lapatinib) growth response in the HER2-overexpressing breast cancer cell line panel. Growth inhibition was determined by methylene blue assay. Shown are the percent inhibitions of cells treated for six days normalized to non-treated cells. The experiment was performed in quadruplicate. Error bars on plots represent +/- standard error (SE). (C) Growth curves of de novo resistant MDA-MB-361 cells treated with different target therapies/regimens for nine days: trastuzumab (T) (10 μg/ml), lapatinib (L) (1 μM), trastuzumab plus lapatinib (L + T), or endocrine therapy, fulvestrant (F) (10 -7 M), untreated (C). Cell numbers were quantified by absorbance at 655 nm after staining with methylene blue. Conditions were repeated in quadruplicate. Significance between groups was determined by multiple comparisons using the Sidak method (* P

    Journal: Breast Cancer Research : BCR

    Article Title: Different mechanisms for resistance to trastuzumab versus lapatinib in HER2- positive breast cancers -- role of estrogen receptor and HER2 reactivation

    doi: 10.1186/bcr3067

    Figure Lengend Snippet: HER2-overexpressing cell lines exhibit distinct responses when treated with potent anti-HER2 therapy . (A) A panel of HER2-overexpressing breast cancer cell lines was treated with lapatinib (1 μM) plus trastuzumab (10 μg/ml) for 48 h and whole-cell extracts were analyzed by immunoblotting with indicated antibodies. (B) Combination therapy (trastuzumab plus lapatinib) growth response in the HER2-overexpressing breast cancer cell line panel. Growth inhibition was determined by methylene blue assay. Shown are the percent inhibitions of cells treated for six days normalized to non-treated cells. The experiment was performed in quadruplicate. Error bars on plots represent +/- standard error (SE). (C) Growth curves of de novo resistant MDA-MB-361 cells treated with different target therapies/regimens for nine days: trastuzumab (T) (10 μg/ml), lapatinib (L) (1 μM), trastuzumab plus lapatinib (L + T), or endocrine therapy, fulvestrant (F) (10 -7 M), untreated (C). Cell numbers were quantified by absorbance at 655 nm after staining with methylene blue. Conditions were repeated in quadruplicate. Significance between groups was determined by multiple comparisons using the Sidak method (* P

    Article Snippet: Lapatinib (Tykerb) was obtained from GlaxoSmithKline (US headquarters in Research Triangle Park, NC, USA) and prepared with dimethyl sulfoxide (DMSO).

    Techniques: Inhibition, Multiple Displacement Amplification, Staining

    Concentration effect curves of (A) single agent cyclopamine and gefitinib and their combinations, (B) single agent cyclopamine and lapatinib and their combinations.

    Journal: Cancer Cell International

    Article Title: Inhibition of androgen-independent prostate cancer cell growth is enhanced by combination therapy targeting Hedgehog and ErbB signalling

    doi: 10.1186/1475-2867-8-3

    Figure Lengend Snippet: Concentration effect curves of (A) single agent cyclopamine and gefitinib and their combinations, (B) single agent cyclopamine and lapatinib and their combinations.

    Article Snippet: Cells were treated with EGF (Sigma), cyclopamine (Sigma), gefitinib (AstraZenica) and lapatinib (Glaxo-Smithkline) as detailed.

    Techniques: Concentration Assay

    Effect of (A) cyclopamine, (B) gefitinib and (C) lapatinib on growth of androgen-independent prostate cancer cells.

    Journal: Cancer Cell International

    Article Title: Inhibition of androgen-independent prostate cancer cell growth is enhanced by combination therapy targeting Hedgehog and ErbB signalling

    doi: 10.1186/1475-2867-8-3

    Figure Lengend Snippet: Effect of (A) cyclopamine, (B) gefitinib and (C) lapatinib on growth of androgen-independent prostate cancer cells.

    Article Snippet: Cells were treated with EGF (Sigma), cyclopamine (Sigma), gefitinib (AstraZenica) and lapatinib (Glaxo-Smithkline) as detailed.

    Techniques:

    (A) Cyclopamine inhibits expression of PTCH and GLI1 RNA in LNCaP C4-2B cells: control cells (lanes 1 and 3); 24 hrs 14 nM cyclopamine treatment (lanes 2 and 4). Relative expression determined by QPCR (error bars show standard deviation). (B) Immunoblot showing inhibitory effect of 168 nM gefitinib on EGFR phosphorylation following EGF treatment in AIPC cells (lane 1: control; lane 2: 24 hr gefitinib; lane 3: 48 hr gefitinib). (C) Immunoblot showing inhibitory effect of 102 nM lapatinib on EGFR phosphorylation following EGF treatment in AIPC cells (lane 1: control; lane 2: 24 hr lapatinib; lane 3: 48 hr lapatinib).

    Journal: Cancer Cell International

    Article Title: Inhibition of androgen-independent prostate cancer cell growth is enhanced by combination therapy targeting Hedgehog and ErbB signalling

    doi: 10.1186/1475-2867-8-3

    Figure Lengend Snippet: (A) Cyclopamine inhibits expression of PTCH and GLI1 RNA in LNCaP C4-2B cells: control cells (lanes 1 and 3); 24 hrs 14 nM cyclopamine treatment (lanes 2 and 4). Relative expression determined by QPCR (error bars show standard deviation). (B) Immunoblot showing inhibitory effect of 168 nM gefitinib on EGFR phosphorylation following EGF treatment in AIPC cells (lane 1: control; lane 2: 24 hr gefitinib; lane 3: 48 hr gefitinib). (C) Immunoblot showing inhibitory effect of 102 nM lapatinib on EGFR phosphorylation following EGF treatment in AIPC cells (lane 1: control; lane 2: 24 hr lapatinib; lane 3: 48 hr lapatinib).

    Article Snippet: Cells were treated with EGF (Sigma), cyclopamine (Sigma), gefitinib (AstraZenica) and lapatinib (Glaxo-Smithkline) as detailed.

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Standard Deviation

    Analysis of cyclopamine and (A-B) gefitinib or (C-D) lapatinib in LNCaP C4-2B cells. (A and C) combination index plot for the drug combinations. (B and D) Isobologram for the combination of gefitinib or (C-D) lapatinib and cyclopamine for effect level (F a = 0.5). Note the combination data points fall on the lower left of the hypotenuse for F a = 0.5 (shown), F a = 0.75 and F a = 0.9 indicating synergism.

    Journal: Cancer Cell International

    Article Title: Inhibition of androgen-independent prostate cancer cell growth is enhanced by combination therapy targeting Hedgehog and ErbB signalling

    doi: 10.1186/1475-2867-8-3

    Figure Lengend Snippet: Analysis of cyclopamine and (A-B) gefitinib or (C-D) lapatinib in LNCaP C4-2B cells. (A and C) combination index plot for the drug combinations. (B and D) Isobologram for the combination of gefitinib or (C-D) lapatinib and cyclopamine for effect level (F a = 0.5). Note the combination data points fall on the lower left of the hypotenuse for F a = 0.5 (shown), F a = 0.75 and F a = 0.9 indicating synergism.

    Article Snippet: Cells were treated with EGF (Sigma), cyclopamine (Sigma), gefitinib (AstraZenica) and lapatinib (Glaxo-Smithkline) as detailed.

    Techniques:

    Lapatinib induces apoptosis in association with downregulation of CIP2A and p-Akt in TNBC cells A. Dose-escalation effects of lapatinib on CIP2A, p-Akt, and caspase 3 cleavage. Cells were exposed to lapatinib at the indicated doses for 48 hours. B. time-dependent analysis of CIP2A, p-Akt, and caspase 3 cleavage. Cells were exposed to lapatinib (10 μM) for 24, 36 and 48 hours. Cell lysates were prepared and assayed for these molecules by western blotting. Data are representative of three independent experiments. Apoptotic cells were determined by flow cytometry (sub-G1 analysis of propidium iodide -stained cells).

    Journal: Oncotarget

    Article Title: Lapatinib inhibits CIP2A/PP2A/p-Akt signaling and induces apoptosis in triple negative breast cancer cells

    doi: 10.18632/oncotarget.7035

    Figure Lengend Snippet: Lapatinib induces apoptosis in association with downregulation of CIP2A and p-Akt in TNBC cells A. Dose-escalation effects of lapatinib on CIP2A, p-Akt, and caspase 3 cleavage. Cells were exposed to lapatinib at the indicated doses for 48 hours. B. time-dependent analysis of CIP2A, p-Akt, and caspase 3 cleavage. Cells were exposed to lapatinib (10 μM) for 24, 36 and 48 hours. Cell lysates were prepared and assayed for these molecules by western blotting. Data are representative of three independent experiments. Apoptotic cells were determined by flow cytometry (sub-G1 analysis of propidium iodide -stained cells).

    Article Snippet: Lapatinib ditosylate (Tykerb) tablets obtained from GlaxoSmithKline (London, UK) were used for in vivo animal experiments.

    Techniques: Western Blot, Flow Cytometry, Cytometry, Staining

    In vivo effect of lapatinib on MDA-MB-468 xenograft nude mice A. Lapatinib treatment decreased the size of MDA-MB-468 tumors. Points , mean ( n = 6); bar , SE. *, P

    Journal: Oncotarget

    Article Title: Lapatinib inhibits CIP2A/PP2A/p-Akt signaling and induces apoptosis in triple negative breast cancer cells

    doi: 10.18632/oncotarget.7035

    Figure Lengend Snippet: In vivo effect of lapatinib on MDA-MB-468 xenograft nude mice A. Lapatinib treatment decreased the size of MDA-MB-468 tumors. Points , mean ( n = 6); bar , SE. *, P

    Article Snippet: Lapatinib ditosylate (Tykerb) tablets obtained from GlaxoSmithKline (London, UK) were used for in vivo animal experiments.

    Techniques: In Vivo, Multiple Displacement Amplification, Mouse Assay

    Lapatinib exerts anti-proliferative and apoptotic-inducing effects in triple-negative breast cancer (TNBC) cells A. Confirmation of HER2 and ER-alpha expression in TNBC cell lines (MDA-MB-231, MDA-MB-468, and HCC-1937). MCF-7 was used as a positive control for ER expression and SK-BR3 was used as a positive control for HER2 expression. B. Left, dose-escalation effects of lapatinib on cell viability; Middle and Right, effects of lapatinib (5 μM), anti-HER2 monoclonal antibody trastuzumab (40 μg/ml), or anti-EGFR monoclonal antibody cetuximab (20 μg/ml) on CIP2A in HER2-positive SK-BR3 cells. HER2, p-HER2, EGFR, p-EGFR, ERK, and p-ERK were assayed to confirm the target effects of these drugs. C. Dose-escalation effects of lapatinib on cell viability in TNBC cells. Cells were exposed to lapatinib at the indicated doses for 72 hours and cell viability was assessed by MTT assay. Points , mean ( n = 3); bars , SD. D. Dose-and time-escalation effects of lapatinib on apoptosis in TNBC cell lines. Cells were exposed to lapatinib at the indicated doses for 24, 48, and 72 hours. Apoptotic cells were determined by flow cytometry (sub-G1 analysis of propidium iodide -stained cells). Columns , mean ( n = 3); bars , SD.

    Journal: Oncotarget

    Article Title: Lapatinib inhibits CIP2A/PP2A/p-Akt signaling and induces apoptosis in triple negative breast cancer cells

    doi: 10.18632/oncotarget.7035

    Figure Lengend Snippet: Lapatinib exerts anti-proliferative and apoptotic-inducing effects in triple-negative breast cancer (TNBC) cells A. Confirmation of HER2 and ER-alpha expression in TNBC cell lines (MDA-MB-231, MDA-MB-468, and HCC-1937). MCF-7 was used as a positive control for ER expression and SK-BR3 was used as a positive control for HER2 expression. B. Left, dose-escalation effects of lapatinib on cell viability; Middle and Right, effects of lapatinib (5 μM), anti-HER2 monoclonal antibody trastuzumab (40 μg/ml), or anti-EGFR monoclonal antibody cetuximab (20 μg/ml) on CIP2A in HER2-positive SK-BR3 cells. HER2, p-HER2, EGFR, p-EGFR, ERK, and p-ERK were assayed to confirm the target effects of these drugs. C. Dose-escalation effects of lapatinib on cell viability in TNBC cells. Cells were exposed to lapatinib at the indicated doses for 72 hours and cell viability was assessed by MTT assay. Points , mean ( n = 3); bars , SD. D. Dose-and time-escalation effects of lapatinib on apoptosis in TNBC cell lines. Cells were exposed to lapatinib at the indicated doses for 24, 48, and 72 hours. Apoptotic cells were determined by flow cytometry (sub-G1 analysis of propidium iodide -stained cells). Columns , mean ( n = 3); bars , SD.

    Article Snippet: Lapatinib ditosylate (Tykerb) tablets obtained from GlaxoSmithKline (London, UK) were used for in vivo animal experiments.

    Techniques: Expressing, Multiple Displacement Amplification, Positive Control, MTT Assay, Flow Cytometry, Cytometry, Staining

    CIP2A/PP2A/p-Akt mediates lapatinib-induced apoptosis in TNBC cells A. ectopic expression of myc-tagged CIP2A reduced the apoptotic effect of lapatinib in MDA-MB-468 cells. B. Analysis of PP2A activity in drug-treated TNBC cells. Cells were treated with DMSO or lapatinib at 10 μM or okadaic acid at 20 nM (as a negative control) or forskolin at 40 μM (as a positive control) for 24 hours. Cell lysates were assayed for PP2A activity. C. Pretreatment of PP2A inhibitor okadaic acid protected cells from lapatinib-induced apoptosis. Cells were pretreated with okadaic acid (20 nM) for 1 hour; then washed and treated with DMSO (control) or lapatinib (10 μM) for 24 hours. Cell lysates were separated and assayed for sub-G1 analysis and western blotting. D. Knockdown of PP2Ac reduced the effects of lapatinib on p-Akt and apoptosis. MDA-MB-468 cells were transfected with siRNA against PP2Ac (catalytic subunit) or control siRNA for 48h, after transfection cells were then treated with lapatinib 5μM for 24 h. Cell lysates were separated and assayed for sub-G1 analysis and western blotting.

    Journal: Oncotarget

    Article Title: Lapatinib inhibits CIP2A/PP2A/p-Akt signaling and induces apoptosis in triple negative breast cancer cells

    doi: 10.18632/oncotarget.7035

    Figure Lengend Snippet: CIP2A/PP2A/p-Akt mediates lapatinib-induced apoptosis in TNBC cells A. ectopic expression of myc-tagged CIP2A reduced the apoptotic effect of lapatinib in MDA-MB-468 cells. B. Analysis of PP2A activity in drug-treated TNBC cells. Cells were treated with DMSO or lapatinib at 10 μM or okadaic acid at 20 nM (as a negative control) or forskolin at 40 μM (as a positive control) for 24 hours. Cell lysates were assayed for PP2A activity. C. Pretreatment of PP2A inhibitor okadaic acid protected cells from lapatinib-induced apoptosis. Cells were pretreated with okadaic acid (20 nM) for 1 hour; then washed and treated with DMSO (control) or lapatinib (10 μM) for 24 hours. Cell lysates were separated and assayed for sub-G1 analysis and western blotting. D. Knockdown of PP2Ac reduced the effects of lapatinib on p-Akt and apoptosis. MDA-MB-468 cells were transfected with siRNA against PP2Ac (catalytic subunit) or control siRNA for 48h, after transfection cells were then treated with lapatinib 5μM for 24 h. Cell lysates were separated and assayed for sub-G1 analysis and western blotting.

    Article Snippet: Lapatinib ditosylate (Tykerb) tablets obtained from GlaxoSmithKline (London, UK) were used for in vivo animal experiments.

    Techniques: Expressing, Multiple Displacement Amplification, Activity Assay, Negative Control, Positive Control, Western Blot, Transfection

    Lapatinib inhibits transcription of CIP2A A. Effect of lapatinib on CIP2A protein degradation. Cells were treated with 100 μg/ml pan-translation inhibitor cyclohexamide (CHX) in the presence (right) or absence (left) of lapatinib (5 μM) for the indicated period of time, then the stability of CIP2A protein in whole-cell lysates was assessed by western blot. The addition of lapatinib did not significantly affect CIP2A degradation. B. Lapatinib inhibited CIP2A transcription. Cells were treated with lapatinib at the indicated doses for 24 hours, after which total RNA was isolated and CIP2A mRNA was assayed by real-time quantitative PCR. Columns , mean ( n = 3); bars , SD. C. Luciferase reporter assay of CIP2A proximal promoter regions upon lapatinib treatment. MDA-MB-468 cells were transfected by Firefly luciferase reporter vectors carrying CIP2A promoters of different lengths as indicated, and Renilla vectors for 24 hours and then treated with 5 μM lapatinib or DMSO for 24 hours. Cell lysates were then assayed for dual luciferase activity as described in Materials and Methods. Columns , mean ( n = 3); bars , SD; *, P

    Journal: Oncotarget

    Article Title: Lapatinib inhibits CIP2A/PP2A/p-Akt signaling and induces apoptosis in triple negative breast cancer cells

    doi: 10.18632/oncotarget.7035

    Figure Lengend Snippet: Lapatinib inhibits transcription of CIP2A A. Effect of lapatinib on CIP2A protein degradation. Cells were treated with 100 μg/ml pan-translation inhibitor cyclohexamide (CHX) in the presence (right) or absence (left) of lapatinib (5 μM) for the indicated period of time, then the stability of CIP2A protein in whole-cell lysates was assessed by western blot. The addition of lapatinib did not significantly affect CIP2A degradation. B. Lapatinib inhibited CIP2A transcription. Cells were treated with lapatinib at the indicated doses for 24 hours, after which total RNA was isolated and CIP2A mRNA was assayed by real-time quantitative PCR. Columns , mean ( n = 3); bars , SD. C. Luciferase reporter assay of CIP2A proximal promoter regions upon lapatinib treatment. MDA-MB-468 cells were transfected by Firefly luciferase reporter vectors carrying CIP2A promoters of different lengths as indicated, and Renilla vectors for 24 hours and then treated with 5 μM lapatinib or DMSO for 24 hours. Cell lysates were then assayed for dual luciferase activity as described in Materials and Methods. Columns , mean ( n = 3); bars , SD; *, P

    Article Snippet: Lapatinib ditosylate (Tykerb) tablets obtained from GlaxoSmithKline (London, UK) were used for in vivo animal experiments.

    Techniques: Western Blot, Isolation, Real-time Polymerase Chain Reaction, Luciferase, Reporter Assay, Multiple Displacement Amplification, Transfection, Activity Assay

    PI3K/Akt signaling is required for VPA-induced upregulation of MICA and MICB in pancreatic cancer cells. (A, B) Quantitative real-time RT-PCR analysis. The VPA-induced upregulation of MICA and MICB mRNA expression were inhibited by the HER2/HER3 inhibitor lapatinib and the PI3K inhibitor LY294002, but not by the ATM/ATR inhibitor caffeine. Data are mean ± SD of a single experiment performed in triplicate, all results were reproducible in three independent experiments. * P

    Journal: BMC Cancer

    Article Title: Valproic acid sensitizes pancreatic cancer cells to natural killer cell-mediated lysis by upregulating MICA and MICB via the PI3K/Akt signaling pathway

    doi: 10.1186/1471-2407-14-370

    Figure Lengend Snippet: PI3K/Akt signaling is required for VPA-induced upregulation of MICA and MICB in pancreatic cancer cells. (A, B) Quantitative real-time RT-PCR analysis. The VPA-induced upregulation of MICA and MICB mRNA expression were inhibited by the HER2/HER3 inhibitor lapatinib and the PI3K inhibitor LY294002, but not by the ATM/ATR inhibitor caffeine. Data are mean ± SD of a single experiment performed in triplicate, all results were reproducible in three independent experiments. * P

    Article Snippet: Lapatinib, LY294002, rabbit polyclonal antibodies against PI3KCA, Akt Rabbit mAb, Phospho-Akt (Ser473) Rabbit mAb, HER3 Rabbit mAb, Phospho-HER3 Rabbit mAb, GAPDH Rabbit mAb, and goat anti-rabbit IgG antibodies conjugated to HRP were purchased from Cell Signaling Technology, Danvers, MA, USA.

    Techniques: Quantitative RT-PCR, Expressing