Journal: PLoS ONE

Article Title: Photo-Activated Psoralen Binds the ErbB2 Catalytic Kinase Domain, Blocking ErbB2 Signaling and Triggering Tumor Cell Apoptosis

doi: 10.1371/journal.pone.0088983

Figure Lengend Snippet: (A) 8MOP interacts with three peptide regions within the ErbB2 catalytic kinase domain. Qualitative peptide identifications within the ErbB2 catalytic kinase domain following LC-MS/MS analysis of a streptavidin pull-down of biotinylated-8MOP bait (see ). The transmembrane domain is indicated (red diamond) and the five C-terminus tyrosine autophosphorylation sites are indicated (p). (B) Non-reducing Western blot analysis of the interaction of 8MOP with ErbB2. BT474 cells were treated with 800dye-8MOP (Promega) or with vehicle (0.01% DMSO) alone served as control for 48 hr and then exposed to UV irradiation (2J) prior to Western blot analysis. The image on the left shows the Western blot for ErbB2 (red). The image on the right shows the same membrane directly scanned for the presence of 800dye-8MOP (green), which overlays the ErbB2 signal. The results are representative of three independent experiments.

Article Snippet: ErbB2+ (BT474; SKBR3) and ErbB2 negative (MCF-7; T47D) human breast cancer cell lines, and the human foreskin fibroblast (HFF) cell line were obtained from the American Type Culture Collection (Manassas, VA).

Techniques: Liquid Chromatography with Mass Spectroscopy, Western Blot, Control, Irradiation, Membrane

Journal: PLoS ONE

Article Title: Photo-Activated Psoralen Binds the ErbB2 Catalytic Kinase Domain, Blocking ErbB2 Signaling and Triggering Tumor Cell Apoptosis

doi: 10.1371/journal.pone.0088983

Figure Lengend Snippet: The growth and viability of BT474 and SKBR3 cells (top bar graphs) after being subjected to the indicated treatment conditions. The combination of PUVA plus neratinib: P <0.0005 (BT474 and SKBR3 cells). Results represent the mean +/− standard error of triplicate samples, and are representative of three independent experiments. (B) Western blot analysis showing steady-state ErbB2, ErbB3, and phospho-Akt (S473) protein levels in BT474 and SKBR3 cells treated according to the indicated treatment conditions. Vehicle alone (0.01% DMSO) served as a control. Steady-state actin protein levels served as a control for equal loading of protein. The results are representative of three independent experiments.

Article Snippet: ErbB2+ (BT474; SKBR3) and ErbB2 negative (MCF-7; T47D) human breast cancer cell lines, and the human foreskin fibroblast (HFF) cell line were obtained from the American Type Culture Collection (Manassas, VA).

Techniques: Western Blot, Control

Journal: PLoS ONE

Article Title: Photo-Activated Psoralen Binds the ErbB2 Catalytic Kinase Domain, Blocking ErbB2 Signaling and Triggering Tumor Cell Apoptosis

doi: 10.1371/journal.pone.0088983

Figure Lengend Snippet: Top bar graph shows the results of the growth assays performed in T47D and stably transfected T47D cell line. T47D cells expressing p85 ErbB2 were pretreated with 5 µM lapatinib or 5 µM 8MOP for 4 hr followed by irradiation in a UV Stratalinker 1800 (Statagene). Cells transfected with empty vector (T47D/Vector), and those treated with vehicle alone (0.01% DMSO) served as controls. The effects of the treatments on cell growth and viability are shown in the bar graph. P<0.0071 (8MOP + UVA irradiation). Results represent the mean +/− standard error of triplicate samples, and are representative of three independent experiments. Steady-state phospho-p85 ErbB2 protein levels (dotted arrow) and phospho-p185 ErbB2 (solid arrow) are shown by Western blot. Actin steady-state protein levels served as a control to ensure for equal loading of protein. Results are representative of three independent experiments.

Article Snippet: ErbB2+ (BT474; SKBR3) and ErbB2 negative (MCF-7; T47D) human breast cancer cell lines, and the human foreskin fibroblast (HFF) cell line were obtained from the American Type Culture Collection (Manassas, VA).

Techniques: Stable Transfection, Transfection, Expressing, Irradiation, Plasmid Preparation, Western Blot, Control

Journal: Endocrinology

Article Title: ATM Is Required for the Prolactin-Induced HSP90-Mediated Increase in Cellular Viability and Clonogenic Growth After DNA Damage.

doi: 10.1210/en.2017-00652

Figure Lengend Snippet: Figure 3. D1-9-G129R-hPRL PRLR antagonist abrogates the PRL-mediated increase in SKBR3 cell viability. (A) Western blot of MCF7 and SKBR3 whole cell extracts probed with the anti-PRLR extracellular domain antibody, 1A2B1. (B) Western blot of nuclear extracts (10 mg/well) of SKBR3 cells starved in serum-free media for 16 hours then treated with 5 mg/mL oPRL or 5 mg/mL D1-9-G129R-hPRL PRLR antagonist (Antag) for 1 hour or left untreated, probed for p-STAT5, STAT5 (rabbit-anti-STAT5), and histone-H3 (HH3) (loading control). (C) A general schematic for treatments used in WST-1 assays including a 24-hour pretreatment, 2 hours of TOPOII poison, followed by a 48-hour recovery period. (D, E, and F) WST-1 assays using SKBR3 cells pretreated or not with 5 mg/mL oPRL 6 D1-9-G129R-hPRL for 24 hours followed by 2 hours of doxorubicin (DOX) treatment, with or without PRL or D1-9-G129R-hPRL, then a 48-hour recovery with or without PRL or D1-9-G129R-hPRL as indicated. (D) The effect of oPRL or PRLR antagonist alone. (E) The effect of DOX 6 PRL and (F) the effect of DOX + Antag 6 PRL. Graphs represents pooled experiments, n = 12. Data were analyzed with a one-way ANOVA followed by Bonferroni posttests. *P , 0.05; **P , 0.01; ***P , 0.001.

Article Snippet: Cell culture and cell lines PRL-responsive SKBR3 (HER2 overexpressing, p53mutant) and MCF7 (estrogen receptor+, p53 wild type) human breast cancer cell lines, obtained and authenticated from American Type Culture Collection, were used within 6 months when revived from frozen storage.

Techniques: Western Blot, Control

Journal: Endocrinology

Article Title: ATM Is Required for the Prolactin-Induced HSP90-Mediated Increase in Cellular Viability and Clonogenic Growth After DNA Damage.

doi: 10.1210/en.2017-00652

Figure Lengend Snippet: Figure 4. HSP90 inhibition abrogates PRL-mediated resistance. (A, B, and C) SKBR3 or (D, E, and F) MCF7 cells were pretreated or not with 5 mg/mL oPRL, 15 nM (SKBR3) or 100 nM (MCF7) HSP90 inhibitor 17-AAG, or a combination of the two for 24 hours followed by 2 hours of doxorubicin (DOX) treatment, 6PRL or 17-AAG, and a 48-hour recovery 6 PRL. (A–C) WST-1 assays of SKBR3 (A) with PRL or 17-AAG alone, (B) with DOX 6 PRL, or (C) with DOX + 17-AAG 6 PRL. (D–F) WST-1 assays of MCF7 (D) with PRL or 17-AAG alone, (E) with DOX 6 PRL, or (F) with DOX + 17-AAG 6 PRL. Graphs represent pooled experiments for SKBR3 n = 12, and for MCF7 n = 18. (G–I) WST-1 assays of MCF7 (G) with oPRL or 800 nM BIIB021 alone, (H) with DOX 6 PRL, or (I) with DOX + BIIB021 6 PRL. n = 18. Data were analyzed with a one-way ANOVA followed by Bonferroni posttests. *P , 0.05; **P , 0.01; ***P , 0.001.

Article Snippet: Cell culture and cell lines PRL-responsive SKBR3 (HER2 overexpressing, p53mutant) and MCF7 (estrogen receptor+, p53 wild type) human breast cancer cell lines, obtained and authenticated from American Type Culture Collection, were used within 6 months when revived from frozen storage.

Techniques: Inhibition

Journal: Endocrinology

Article Title: ATM Is Required for the Prolactin-Induced HSP90-Mediated Increase in Cellular Viability and Clonogenic Growth After DNA Damage.

doi: 10.1210/en.2017-00652

Figure Lengend Snippet: Figure 6. JAK2 inhibition abrogates the PRL increased cell viability. (A) Western blot of JAK2 inhibitor dose response. MCF7 cells were treated with twofold increasing concentrations of G6 for 12 hours. p-STAT5 levels were determined (mouse anti-p-STAT5) with total STAT5 and histone H3 (HH3) loading controls. Results are representative of two independent experimental replicates. (B) Dose-response curves of JAK2 inhibitor G6. MCF7 cells were treated with G6 at indicated time points and concentrations. The cell viability was determined with WST-1 cell viability assay. WST-1 assays of (C–E) MCF7 or (F–H) SKBR3 cells pretreated with 25 ng/mL hPRL for 24 hours or 25 mM JAK2 inhibitor G6 for 12 hours, followed by

Article Snippet: Cell culture and cell lines PRL-responsive SKBR3 (HER2 overexpressing, p53mutant) and MCF7 (estrogen receptor+, p53 wild type) human breast cancer cell lines, obtained and authenticated from American Type Culture Collection, were used within 6 months when revived from frozen storage.

Techniques: Inhibition, Western Blot, Viability Assay

Journal: Endocrinology

Article Title: ATM Is Required for the Prolactin-Induced HSP90-Mediated Increase in Cellular Viability and Clonogenic Growth After DNA Damage.

doi: 10.1210/en.2017-00652

Figure Lengend Snippet: Figure 7. HSP90 inhibition decreases ATM and p-ATM protein but not its mRNA. (A) MCF7 cells were treated or not with threefold increasing concentrations of 17-AAG for 24 hours followed by 2 hours of doxorubicin (DOX) treatment and recovery. Proteins (30 mg per well) were resolved on SDS-PAGE gel, and blot was probed for p-ATM, ATM, HSP90a, and GRB2. Gel is representative of two experiments. Fold difference of p-ATM with DOX treatment was calculated, in ImageJ, compared with untreated cells (set to 1). Fold difference of (B) total ATM and (C) p- ATM levels with 17-AAG and DOX treatment were calculated compared with DOX alone (set to 1). Representative blot of two experiments. (D) MCF7 cells were pretreated with oPRL (5 mg/mL) or 17-AAG for 24 hours, followed by 2 hours DOX treatment (0.2 mM). ATM expression was detected by qPCR, and the results were normalized to YWHAZ control [mean 6 standard error of the mean (SEM); n = 5]. The DDCt method was used to analyze the relative changes in gene. The letter “a” above SEM bars denotes that there were no statistically significant differences (one-way ANOVA followed by Bonferroni test, P , 0.05). (E) SKBR3 cells as above, with (F) ATM and (G) p-ATM band intensities quantified by ImageJ. Representative blot of two experiments.

Article Snippet: Cell culture and cell lines PRL-responsive SKBR3 (HER2 overexpressing, p53mutant) and MCF7 (estrogen receptor+, p53 wild type) human breast cancer cell lines, obtained and authenticated from American Type Culture Collection, were used within 6 months when revived from frozen storage.

Techniques: Inhibition, SDS Page, Expressing, Control

Journal: Endocrinology

Article Title: ATM Is Required for the Prolactin-Induced HSP90-Mediated Increase in Cellular Viability and Clonogenic Growth After DNA Damage.

doi: 10.1210/en.2017-00652

Figure Lengend Snippet: Figure 11. ATM is needed for PRL-induced increase in cell viability after DNA damage in 3D culture. Cells were pretreated with 25 ng/mL hPRL or vehicle for 24 hours followed by 2 hours of 1 mM doxorubicin (DOX) treatment (or vehicle) with or without PRL. Cells were transferred to collagen 3D culture and allowed to recover for 48 hours. Cell viability was determined with WST-1 assay, and the treatments were normalized to vehicle controls: (A) SKBR3 or (B) MCF7. For ATM knockdown experiments, (C and E) SKBR3 and (D and F) MCF7 cells were transfected with (C and D) siNT or (E and F) siATM, followed by 24 hours of 25 ng/mL hPRL and 2 hours of DOX treatment (3 mM for SKBR3, 2 mM for MCF7 cells). Cells were transferred to collagen gels to allow recovery for 96 hours (SKBR3) or 48 hours (MCF7). Bars in the graphs represent six independent experiments. Statistically significant analysis by ANOVA: *P , 0.05; **P , 0.01; ***P , 0.001.

Article Snippet: Cell culture and cell lines PRL-responsive SKBR3 (HER2 overexpressing, p53mutant) and MCF7 (estrogen receptor+, p53 wild type) human breast cancer cell lines, obtained and authenticated from American Type Culture Collection, were used within 6 months when revived from frozen storage.

Techniques: WST-1 Assay, Knockdown, Transfection

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: New HER2-negative breast cancer subtype responsive to anti-HER2 therapy identified

doi: 10.1007/s00432-020-03144-7

Figure Lengend Snippet: Percent inhibition of NRG1b CELx signal for HER2+ cell lines versus HER2−/HSFs+ patient samples . Inhibition of NRG1-driven HER2 signaling by pertuzumab (10 mg/mL) or trastuzumab (10 mg/mL) alone or in combination in the CELx test. Percentages shown represent the average of four HER2+ HSFs+ cell lines (SKBR3, EFM192A, HCC1569, and ZR75-30) or 5 HER2−/HSFs+ primary tumor samples (C978, C133, C264, C309, and C371), performed with 2 technical replicates per biological sample

Article Snippet: Human breast cancer cell lines used in this study included SKBr3 (HER2+ /HSFs+), BT474 (HER2+ /HSFs−), BT483 (ER-/HER2−/HSFs+), ZR75-30 (HER2+ /HSFs+), HCC1569 (HER2+ /HSFs−), HCC1954 (ER-/HER2+ /HSFs−), HCC202 (HER2+ /HSFs+), MDA-MB361 (HER2+ /HSFs−), AU565 (HER2+ /HSFs−) (all from American Tissue Type Collection; ATCC, Manassas, VA), and EFM192A (HER2+ /HSFs+) (from Leibniz Institute DSMZ, Germany).

Techniques: Inhibition

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: New HER2-negative breast cancer subtype responsive to anti-HER2 therapy identified

doi: 10.1007/s00432-020-03144-7

Figure Lengend Snippet: Comparison of inhibition of NRG1-driven HER2 signaling by HER2 targeted therapeutics. Pertuzumab (10 μg/mL), lapatinib (200 nM), afatinib (120 nM), or neratinib (500 nM) inhibition of NRG1-driven HER2 signaling is shown as the average of 9 HER2+ cell lines (SKBR3, EFM192A, ZR75-30, HCC202, HCC1954, HCC1569, MDA-MB361, BT474, AU565) or 7 HER2−/HER2s+ primary tumors (R69, R20, R160, R82, R95, R25, R71) performed with 2 technical replicates per biological sample. Single dose concentrations were selected for relative comparator purposes only where a maximal amount of the signaling was inhibited in at least one of the samples. Variances from 100% inhibition likely arise from ligand generated signaling that was not directly related to HER activity or where a sample had some level of drug paradoxical response (Claus et al. )

Article Snippet: Human breast cancer cell lines used in this study included SKBr3 (HER2+ /HSFs+), BT474 (HER2+ /HSFs−), BT483 (ER-/HER2−/HSFs+), ZR75-30 (HER2+ /HSFs+), HCC1569 (HER2+ /HSFs−), HCC1954 (ER-/HER2+ /HSFs−), HCC202 (HER2+ /HSFs+), MDA-MB361 (HER2+ /HSFs−), AU565 (HER2+ /HSFs−) (all from American Tissue Type Collection; ATCC, Manassas, VA), and EFM192A (HER2+ /HSFs+) (from Leibniz Institute DSMZ, Germany).

Techniques: Comparison, Inhibition, Generated, Activity Assay

Journal: Medicina

Article Title: Eribulin Mesylate Improves Cisplatin-Induced Cytotoxicity of Triple-Negative Breast Cancer by Extracellular Signal-Regulated Kinase 1/2 Activation

doi: 10.3390/medicina58040547

Figure Lengend Snippet: Effect of cisplatin and eribulin on the viability of HCC38, MDA-MB-231, and SKBR3 breast cancer cells. Cells were treated with cisplatin (3, 15, 30, and 60 µM) ( A ) or eribulin (12, 30, 60, and 120 µM) ( B ) for 24 h. Cell viability was measured using the Cell Counting Kit-8 assay. Each assay was performed in triplicate. Data are presented as the mean ± standard deviation; * p < 0.05 vs. untreated control.

Article Snippet: HCC38 (ATCC CRL-2314 TM ; ER-, PR-, HER2-), MDA-MB-231 (ATCC HTB-26 TM ; ER-, PR-, HER2-), and SKBR3 (ATCC HTB-30 TM ; HER2+) cells were acquired from the American Type Culture Collection (ATCC) (Manassas, VA, USA).

Techniques: Cell Counting, Standard Deviation, Control

Journal: Medicina

Article Title: Eribulin Mesylate Improves Cisplatin-Induced Cytotoxicity of Triple-Negative Breast Cancer by Extracellular Signal-Regulated Kinase 1/2 Activation

doi: 10.3390/medicina58040547

Figure Lengend Snippet: Apoptotic effect of cisplatin or eribulin on PARP cleavage in HCC38, MDA-MB-231, and SKBR3 breast cancer cells. Cells were treated with cisplatin (1.5, 3, 15, 30, and 60 µM) ( A ) or eribulin (12 and 60 µM) ( B ) for 24 h. Cleavage of the caspase substrate PARP in the presence of cisplatin or eribulin is shown. Cells were harvested after 24-h cisplatin or eribulin treatment. PARP cleavage was assessed by Western blot analysis using an anti-PARP antibody. β-Actin was used as the internal control. Data are presented as the mean ± standard deviation; * p < 0.05 vs. untreated control.

Article Snippet: HCC38 (ATCC CRL-2314 TM ; ER-, PR-, HER2-), MDA-MB-231 (ATCC HTB-26 TM ; ER-, PR-, HER2-), and SKBR3 (ATCC HTB-30 TM ; HER2+) cells were acquired from the American Type Culture Collection (ATCC) (Manassas, VA, USA).

Techniques: Western Blot, Control, Standard Deviation

Journal: Medicina

Article Title: Eribulin Mesylate Improves Cisplatin-Induced Cytotoxicity of Triple-Negative Breast Cancer by Extracellular Signal-Regulated Kinase 1/2 Activation

doi: 10.3390/medicina58040547

Figure Lengend Snippet: Combined effect of cisplatin and eribulin on HCC38, MDA-MB-231, and SKBR3 breast cancer cells. Cells were treated with cisplatin (1.5, 3, 15, and 30 µM) or a combination of cisplatin and eribulin (60 µM). After 24 h of exposure, cell viability was measured using a Cell Counting Kit-8 assay. Each assay was performed in triplicate. Data are presented as the mean ± standard deviation; * p < 0.05 vs. cisplatin-only treatment.

Article Snippet: HCC38 (ATCC CRL-2314 TM ; ER-, PR-, HER2-), MDA-MB-231 (ATCC HTB-26 TM ; ER-, PR-, HER2-), and SKBR3 (ATCC HTB-30 TM ; HER2+) cells were acquired from the American Type Culture Collection (ATCC) (Manassas, VA, USA).

Techniques: Cell Counting, Standard Deviation

Journal: Medicina

Article Title: Eribulin Mesylate Improves Cisplatin-Induced Cytotoxicity of Triple-Negative Breast Cancer by Extracellular Signal-Regulated Kinase 1/2 Activation

doi: 10.3390/medicina58040547

Figure Lengend Snippet: Combination index (CI) values for cisplatin and eribulin combination for breast cancer cell lines.

Article Snippet: HCC38 (ATCC CRL-2314 TM ; ER-, PR-, HER2-), MDA-MB-231 (ATCC HTB-26 TM ; ER-, PR-, HER2-), and SKBR3 (ATCC HTB-30 TM ; HER2+) cells were acquired from the American Type Culture Collection (ATCC) (Manassas, VA, USA).

Techniques:

Journal: Medicina

Article Title: Eribulin Mesylate Improves Cisplatin-Induced Cytotoxicity of Triple-Negative Breast Cancer by Extracellular Signal-Regulated Kinase 1/2 Activation

doi: 10.3390/medicina58040547

Figure Lengend Snippet: Eribulin enhances the apoptotic effect of cisplatin on TNBC cells. ( A ) HCC38, MDA-MB-231, and SKBR3 cells were exposed to cisplatin (15 and 30 µM) or eribulin (60 µM) individually or in combination for 24 h. Cell lysates were collected and subjected to Western blot analysis with the anti-PARP antibody. β-Actin was used as the internal control; ** p < 0.01 vs. cisplatin-only (15 µM) treatment in cleaved PARP, †† p < 0.01 vs. cisplatin-only (30 µM) treatment in cleaved PARP, ‡‡ p < 0.01 vs. eribulin-only (60 µM) treatment in cleaved PARP. (B) Cells were treated with cisplatin alone, eribulin alone (60 µM), or a combination of cisplatin and eribulin for 24 h. The absorbance of pNA (p-nitroaniline) at 405 nm was measured. Results are presented as the mean ± standard deviation from three independent studies; * p < 0.05 vs. cisplatin-only (30 µM) treatment, † p < 0.05, †† p < 0.01 vs. eribulin-only (60 µM) treatment.

Article Snippet: HCC38 (ATCC CRL-2314 TM ; ER-, PR-, HER2-), MDA-MB-231 (ATCC HTB-26 TM ; ER-, PR-, HER2-), and SKBR3 (ATCC HTB-30 TM ; HER2+) cells were acquired from the American Type Culture Collection (ATCC) (Manassas, VA, USA).

Techniques: Western Blot, Control, Standard Deviation

Journal: Medicina

Article Title: Eribulin Mesylate Improves Cisplatin-Induced Cytotoxicity of Triple-Negative Breast Cancer by Extracellular Signal-Regulated Kinase 1/2 Activation

doi: 10.3390/medicina58040547

Figure Lengend Snippet: Eribulin and cisplatin in combination synergistically activated ERK activation in TNBC cells. HCC38, MDA-MB-231, and SKBR3 cells were exposed to cisplatin (3, 15, 30, and 60 µM) ( A ) or eribulin (12 and 60 µM) ( B ) for 24 h. Cell lysates were collected and subjected to Western blot analysis with anti-phospho-ERK1/2 and anti-ERK1/2 antibodies. β-Actin was used as the internal control. Data are presented as the mean ± standard deviation; * p < 0.05 vs. untreated control. ( C ) Cells were exposed to cisplatin (30 µM) or eribulin (60 µM) individually or in combination for 24 h. Levels of phosphorylated ERK1/2 and total-ERK1/2 were assessed by Western blot analysis. β-Actin was used as the internal control; * p < 0.05 vs. cisplatin-only (30 µM) treatment, † p < 0.05, †† p < 0.01 vs. eribulin-only (60 µM) treatment.

Article Snippet: HCC38 (ATCC CRL-2314 TM ; ER-, PR-, HER2-), MDA-MB-231 (ATCC HTB-26 TM ; ER-, PR-, HER2-), and SKBR3 (ATCC HTB-30 TM ; HER2+) cells were acquired from the American Type Culture Collection (ATCC) (Manassas, VA, USA).

Techniques: Activation Assay, Western Blot, Control, Standard Deviation

Journal: Medicina

Article Title: Eribulin Mesylate Improves Cisplatin-Induced Cytotoxicity of Triple-Negative Breast Cancer by Extracellular Signal-Regulated Kinase 1/2 Activation

doi: 10.3390/medicina58040547

Figure Lengend Snippet: Phosphorylation of ERK1/2 in HCC38, MDA-MB-231, and SKBR3 cells treated with PD98059 and the combination of cisplatin and eribulin. ( A ) Cells were first treated with 25 µM PD98059 for 2 h and then cultured with cisplatin (30 µM) and eribulin (60 µM) for 24 h. Total ERK1/2 and phospho-ERK1/2 levels were analyzed by Western blot analysis. β-Actin was used as the internal control; * p < 0.05 vs. cisplatin (30 µM) + eribulin (60 µM). ( B ) Cell lysates were collected and subjected to Western blot analysis with the anti-PARP antibody. β-Actin was used as the internal control; ** p < 0.01 vs. cisplatin (30 µM) + eribulin (60 µM) treatment in cleaved PARP. ( C ) After 24 h exposure, cell viability was measured using a Cell Counting Kit-8 assay. Each assay was performed in triplicate. Data are presented as the mean ± standard deviation; * p < 0.05 vs. cisplatin (30 µM) + eribulin (60 µM).

Article Snippet: HCC38 (ATCC CRL-2314 TM ; ER-, PR-, HER2-), MDA-MB-231 (ATCC HTB-26 TM ; ER-, PR-, HER2-), and SKBR3 (ATCC HTB-30 TM ; HER2+) cells were acquired from the American Type Culture Collection (ATCC) (Manassas, VA, USA).

Techniques: Phospho-proteomics, Cell Culture, Western Blot, Control, Cell Counting, Standard Deviation

Journal: Cell Death & Disease

Article Title: Intracellular cartilage oligomeric matrix protein augments breast cancer resistance to chemotherapy

doi: 10.1038/s41419-024-06872-7

Figure Lengend Snippet: Cyquant survival assay was performed using ( A – E ) BT-20 and ( F – J ) MDA-MB-231 cells expressing COMP and mock control treated with doxorubicin, epirubicin, paclitaxel, docetaxel, and 5-fluoracil, respectively, at the indicated concentrations. K , L Cyquant survival assay was performed in MCF-7 cells expressing COMP and respective mock control cells treated with tamoxifen and fulvestrant. M , N Cyquant survival assay was conducted in HER2-positive SKBR3 cells expressing COMP and mock control cells treated with pertuzumab and trastuzumab. Late apoptotic and live cells were determined by flow cytometry in both ( O , P ) BT-20 and ( Q , R ) MDA-MB 231 cells following anti-cancer chemotherapeutic agents treatment. Data are representative of at least three independent experiments. Statistical significances were determined by repeat measurements two-way ANOVA with Sidak post-hoc test. *p < 0.05; **p < 0.01; ***p < 0.001 and ****p < 0.0001.

Article Snippet: The HER2 positive SKBR3 cells expressing COMP and mock were treated with pertuzumab (Selleckchem) and trastuzumab (Selleckchem) for 72 h at concentrations showed in the figures.

Techniques: CyQUANT Assay, Clonogenic Cell Survival Assay, Expressing, Control, Flow Cytometry

Journal: Frontiers in Oncology

Article Title: The effect of the alpha-specific PI3K inhibitor alpelisib combined with anti-HER2 therapy in HER2+/PIK3CA mutant breast cancer

doi: 10.3389/fonc.2023.1108242

Figure Lengend Snippet: Cell growth inhibition in vitro of HER2+ breast cancer cell lines treated with monoclonal anti-HER2 antibodies. Cells were plated in 24-well plates and treated with vehicle (DMSO), 10 μg/ml trastuzumab, or 10 μg/ml trastuzumab + 10 μg/ml pertuzumab. Cell growth was assessed after 6 days. Data were normalized to vehicle. Data represent means ± SEM. *p-value <0.05, **p-value <0.01; ***p-value <0.001; ****p-value <0.0001; ns, not significant according to multiple t test with Bonferroni correction.

Article Snippet: HER2+ breast cancer cell lines SKBR3 (ATCC ® HTB-30TM), BT474 (ATCC ® HTB-20TM), and HCC-1954 (ATCC ® CRL-2338TM) were obtained from the American Type Culture Collection (ATCC).

Techniques: Inhibition, In Vitro

Journal: Frontiers in Oncology

Article Title: The effect of the alpha-specific PI3K inhibitor alpelisib combined with anti-HER2 therapy in HER2+/PIK3CA mutant breast cancer

doi: 10.3389/fonc.2023.1108242

Figure Lengend Snippet: Responses of HER2+ breast cancer cell lines to the alpha-specific PI3K inhibitor alpelisib. (A) , Cells were plated in 24-well plates and treated with increasing doses of alpelisib. Cell growth was assessed after 6 days. Data were analyzed by GraphPad Prism (version 9.0) to generate drug response curves and relative IC50 values using the log (inhibitor) versus response-variable slope model (bars, SEM) with normalization of data defining the biggest number in each dataset as 100% and the smallest number in the same dataset as 0%. (B) , Estimated IC50 values of alpelisib in HER2 + breast cancer cell lines. CI, confidence of interval.

Article Snippet: HER2+ breast cancer cell lines SKBR3 (ATCC ® HTB-30TM), BT474 (ATCC ® HTB-20TM), and HCC-1954 (ATCC ® CRL-2338TM) were obtained from the American Type Culture Collection (ATCC).

Techniques:

Journal: Frontiers in Oncology

Article Title: The effect of the alpha-specific PI3K inhibitor alpelisib combined with anti-HER2 therapy in HER2+/PIK3CA mutant breast cancer

doi: 10.3389/fonc.2023.1108242

Figure Lengend Snippet: The alpha-specific PI3K inhibitor alpelisib increases the efficacy of anti-HER2 therapy in HER2+ breast cancer cell lines. (A) , Cells were plated in 24-multiwell and treated with vehicle (DMSO), 10μg/ml trastuzumab, 10μg/ml trastuzumab + 10μg/ml pertuzumab, 1μM alpelisib, 1μM alpelisib + 10μg/ml trastuzumab, 1μM alpelisib + 10μg/ml trastuzumab + 10μg/ml pertuzumab. Cell growth was assessed after 6 days. Data were normalized to vehicle. Data represent means ± SEM. *p-value <0.05; ***p-value <0.001; ****p-value <0.0001 according to multiple t test with Bonferroni correction. (B) , Western blot analysis of total and phosphorylated proteins of the HER2/PI3K pathway signaling. Cells were plated in 6-multiwell and treated with vehicle (DMSO), 10μg/ml trastuzumab (T), 10μg/ml trastuzumab + 10μg/ml pertuzumab (T+P), 1μM alpelisib (A), 1μM alpelisib + 10μg/ml trastuzumab (A+T), 1μM alpelisib + 10μg/ml trastuzumab + 10μg/ml pertuzumab (A+T+P).

Article Snippet: HER2+ breast cancer cell lines SKBR3 (ATCC ® HTB-30TM), BT474 (ATCC ® HTB-20TM), and HCC-1954 (ATCC ® CRL-2338TM) were obtained from the American Type Culture Collection (ATCC).

Techniques: Western Blot

Journal: Frontiers in Oncology

Article Title: The effect of the alpha-specific PI3K inhibitor alpelisib combined with anti-HER2 therapy in HER2+/PIK3CA mutant breast cancer

doi: 10.3389/fonc.2023.1108242

Figure Lengend Snippet: Pharmacological synergy between alpelisib and trastuzumab in HER2+/ PIK3CA mutant breast cancer cell lines. HCC1954 and KPL4 cells were seeded in 96 well plate treated with increasing concentrations of each drug alone or in combination (up to 10μM alpelisib and 100μg/ml trastuzumab) for 7 days. Combination indices were determined using the Chou-Talalay test by CompuSyn software. Numbers inside each box indicate the ratio of viable treated cells to untreated cells, from three independent experiments. Synergy is defined as combination index (CI)<1, an additive effect as CI=1, and antagonism effect as CI>1.

Article Snippet: HER2+ breast cancer cell lines SKBR3 (ATCC ® HTB-30TM), BT474 (ATCC ® HTB-20TM), and HCC-1954 (ATCC ® CRL-2338TM) were obtained from the American Type Culture Collection (ATCC).

Techniques: Mutagenesis, Software

Journal: Frontiers in Oncology

Article Title: The effect of the alpha-specific PI3K inhibitor alpelisib combined with anti-HER2 therapy in HER2+/PIK3CA mutant breast cancer

doi: 10.3389/fonc.2023.1108242

Figure Lengend Snippet: Venn diagram of significantly up-regulated genes in HER2+/ PIK3CA mutant alpelisib-resistant (AR) and alpelisib + trastuzumab-resistant (ATR) derivatives.

Article Snippet: HER2+ breast cancer cell lines SKBR3 (ATCC ® HTB-30TM), BT474 (ATCC ® HTB-20TM), and HCC-1954 (ATCC ® CRL-2338TM) were obtained from the American Type Culture Collection (ATCC).

Techniques: Mutagenesis

Journal: Frontiers in Oncology

Article Title: The effect of the alpha-specific PI3K inhibitor alpelisib combined with anti-HER2 therapy in HER2+/PIK3CA mutant breast cancer

doi: 10.3389/fonc.2023.1108242

Figure Lengend Snippet: Kaplan-Meier curves showing relapse-free survival (RFS) of HER2+ breast cancer patients. Log-rank test was used for comparing two groups with high (red) and low (black) expression of ARL4A , SNRPG , ODC1 , AKR1C1 , TRIM16L , AKR1C2 , GLB1L2 , and BDH1 genes. Number of patients at risk are indicated below each panel. HR, hazard ratio.

Article Snippet: HER2+ breast cancer cell lines SKBR3 (ATCC ® HTB-30TM), BT474 (ATCC ® HTB-20TM), and HCC-1954 (ATCC ® CRL-2338TM) were obtained from the American Type Culture Collection (ATCC).

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: The effect of the alpha-specific PI3K inhibitor alpelisib combined with anti-HER2 therapy in HER2+/PIK3CA mutant breast cancer

doi: 10.3389/fonc.2023.1108242

Figure Lengend Snippet: Effect of AKR1C1 siRNA knockdown in parental and alpelisib-resistant HER2+/ PIK3CA mutant breast cancer cell lines. (A) , HCC1954 and KPL4 parental and alpelisib-resistant (AR) cell lines were transfected with siRNA targeting the AKR1C1 or siRNA control (si-Cnt). Culture medium was replaced the next day with regular medium or alpelisib-containing medium, and replaced again at 4 days. Cell growth was assessed at day 6. Data were normalized to si-Cnt transfection. Data represent means ± SEM. *p-value <0.05; ****p-value <0.0001 according to multiple t test with Bonferroni correction. (B) , Protein levels of AKR1C1 in HER2+/PIK3CA mutant breast cancer cell lines transfected with siRNA targeting siRNA control or against AKR1C1 were tested by Western blot.

Article Snippet: HER2+ breast cancer cell lines SKBR3 (ATCC ® HTB-30TM), BT474 (ATCC ® HTB-20TM), and HCC-1954 (ATCC ® CRL-2338TM) were obtained from the American Type Culture Collection (ATCC).

Techniques: Knockdown, Mutagenesis, Transfection, Control, Western Blot

Journal: American Journal of Translational Research

Article Title: Maytansine-loaded star-shaped folate-core PLA-TPGS nanoparticles enhancing anticancer activity

doi:

Figure Lengend Snippet: Fluorescence microscopy images images of SKBR3 after 4 h of incubation with the coumarin 6-loaded star FA-PLA-TPGS nanoparticles (coumarin 6-NPs). The coumarin 6-loaded nanoparticles were green (×100). A. SKBR3 cells incubated with coumarin 6-NPs for 12 hours; B. SKBR3 cells incubated with coumarin 6-NPs for 24 hours; C. SKBR3 cells incubated with coumarin 6-NPs for 36 hours.

Article Snippet: Human breast adenocarcinoma cell line SKBR3 (HER2-over expression) was obtained from American Type Culture Collection (ATCC; Rockville, MD, USA).

Techniques: Fluorescence, Microscopy, Incubation

Journal: American Journal of Translational Research

Article Title: Maytansine-loaded star-shaped folate-core PLA-TPGS nanoparticles enhancing anticancer activity

doi:

Figure Lengend Snippet: Growth inhibitory effects of maytansine and maytansine-NPs on SKBR3 cell lines in vitro. The effects of maytansine and maytansine-NPs (equivalent in maytansine) was tested at a concentrations 0.001, 0.01, 0.1, 1.0 and 10.0 nmol/L after treatment 48 h and 72 h, respectively. Both drug inhibited the growth of SKBR3 cells in a dose-dependent manner, but the effect on maytansine-NPs had no difference with that of maytansine in the concentration range of 0.01-10.0 nmol/L (p > 0.05).

Article Snippet: Human breast adenocarcinoma cell line SKBR3 (HER2-over expression) was obtained from American Type Culture Collection (ATCC; Rockville, MD, USA).

Techniques: In Vitro, Concentration Assay