Journal: mAbs

Article Title: Structural and functional characterization of MBS301, an afucosylated bispecific anti-HER2 antibody

doi: 10.1080/19420862.2018.1486946

Figure Lengend Snippet: ADCC activity evaluation of anti-HER2 antibodies on four target cancer cells.

Article Snippet: Human breast cancer cell line BT-474 (HTB-20), MDA-MB-175VII (HTB-25), SK-BR-3(HTB-30) and HCC1419(CRL-2326) are HER2-positive cell lines, which were obtained from the ATCC, and HCC1419 is trastuzumab-resistant human breast cancer cell line.

Techniques: Activity Assay

Journal: International Journal of Nanomedicine

Article Title: Polymer–lipid hybrid anti-HER2 nanoparticles for targeted salinomycin delivery to HER2-positive breast cancer stem cells and cancer cells

doi: 10.2147/IJN.S144184

Figure Lengend Snippet: Expression of HER2 in breast cancer cells, analyzed by flow cytometry. Notes: Representative fluorescence-activated cell sorting analysis of ( A ) MDA-MB-361 and ( B ) BT-474 cells tested by ALDEFLUOR assay. In the right image (ALDH+ DEAB), cells incubated with ALDH substrate (BAAA) and the specific inhibitor of ALDH, DEAB, were used to establish the baseline fluorescence of these cells (R1) and to define the ALDEFLUOR-positive region (R2). ( C ) Percentage of HER2 positively stained cells in breast cancer cells. ( D ) HER2 mean fluorescence intensity in breast cancer cells. Data are expressed as mean ± SD (n=3). ** P <0.01. Abbreviations: ALDH, aldehyde dehydrogenase; BAAA, BODIPY-aminoacetaldehyde; DEAB, diethylaminobenzaldehyde.

Article Snippet: Human breast cancer cell lines BT-474 and MDA-MB-361 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Flow Cytometry, Fluorescence, FACS, Incubation, Staining

Journal: International Journal of Nanomedicine

Article Title: Polymer–lipid hybrid anti-HER2 nanoparticles for targeted salinomycin delivery to HER2-positive breast cancer stem cells and cancer cells

doi: 10.2147/IJN.S144184

Figure Lengend Snippet: Cell proliferation assay. Notes: ALDH+ and ALDH− breast cancer cells were seeded in 96-well plates with a density of 1×10 4 cells per well overnight. The cells were incubated with the nanoparticles or salinomycin for 72 h, and cell viability was evaluated using the CCK-8 assay. ( A ) MDA-MB-361 ALDH+, ( B ) MDA-MB-361 ALDH−, ( C ) BT-474 ALDH+, and ( D ) BT-474 ALDH−. Data are expressed as mean ± SD (n=3). Abbreviations: ALDH, aldehyde dehydrogenase; CCK-8, Cell Counting Kit-8; Sali-NP, salinomycin-loaded polymer-lipid nanoparticles; Sali-NP-HER2, salinomycin-loaded polymer–lipid hybrid anti-HER2 nanoparticles.

Article Snippet: Human breast cancer cell lines BT-474 and MDA-MB-361 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Proliferation Assay, Incubation, CCK-8 Assay, Cell Counting

Journal: International Journal of Nanomedicine

Article Title: Polymer–lipid hybrid anti-HER2 nanoparticles for targeted salinomycin delivery to HER2-positive breast cancer stem cells and cancer cells

doi: 10.2147/IJN.S144184

Figure Lengend Snippet: The IC 50 values of salinomycin and nanoparticles in breast cancer cells

Article Snippet: Human breast cancer cell lines BT-474 and MDA-MB-361 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques:

Journal: International Journal of Nanomedicine

Article Title: Polymer–lipid hybrid anti-HER2 nanoparticles for targeted salinomycin delivery to HER2-positive breast cancer stem cells and cancer cells

doi: 10.2147/IJN.S144184

Figure Lengend Snippet: Effect of treatments on the percentage of CSCs. Notes: Effect of the treatments on the proportion of CSCs in breast cancer cells, as reflected by the tumorsphere formation rate ( A and B ) and the proportion of ALDH+ cells ( E and F ). Representative images of tumorspheres formed by ( C ) MDA-MB-361 cells and ( D ) BT-474 cells are shown. The rate of tumorsphere formation is defined as the number of tumorspheres formed in 7 days in the treatment group divided by the number of tumorspheres formed in 7 days in the untreated group; the rate of tumorsphere formation in the untreated group is used as a control and defined as 100%. The two groups were compared by one-way ANOVA with the Newman–Keuls method. Data are expressed as mean ± SD (n=6). * P <0.05; ** P <0.01; *** P <0.001. Abbreviations: ANOVA, analysis of variance; ALDH, aldehyde dehydrogenase; CSCs, cancer stem cells; Sali-NP, salinomycin-loaded polymer–lipid nanoparticles; NP-HER2, polymer–lipid anti-HER2 nanoparticles; Sali-NP-HER2, salinomycin-loaded polymer–lipid hybrid anti-HER2 nanoparticles.

Article Snippet: Human breast cancer cell lines BT-474 and MDA-MB-361 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques:

Journal: International Journal of Nanomedicine

Article Title: Polymer–lipid hybrid anti-HER2 nanoparticles for targeted salinomycin delivery to HER2-positive breast cancer stem cells and cancer cells

doi: 10.2147/IJN.S144184

Figure Lengend Snippet: In vivo antitumor assay in mice bearing subcutaneous BT-474 tumors. Notes: BT-474 tumors reached ~50 mm 3 in size by day 10. From day 10, mice were injected with nanoparticles (7.5 mg salinomycin/kg, i.v.) through the tail vein, and salinomycin (7.5 mg salinomycin/kg) dissolved in ethanol was administered by intraperitoneal injection. Therapy was given nine times on alternate days (indicated by arrows), and tumor volume was calculated. ( A ) Tumor growth curve. ( B ) Excised tumors. ( C ) The excised tumors were weighed at the end point. On day 28, the effect of the drug treatments on the CSC proportion in BT-474 tumors in vivo was evaluated by ( D ) the rate of tumorsphere formation and ( E ) the proportion of ALDH+ cells from the excised tumors. ( F ) Representative images of tumorspheres from ( D ) are shown. The two groups were compared by one-way ANOVA with the Newman–Keuls method. Data are expressed as mean ± SD (n=8). * P <0.05; ** P <0.01; *** P <0.001. Abbreviations: ANOVA, analysis of variance; ALDH, aldehyde dehydrogenase; CSC, cancer stem cell; i.v., intravenous; Sali-NP, salinomycin-loaded polymer–lipid nanoparticles; NP-HER2, polymer–lipid anti-HER2 nanoparticles; Sali-NP-HER2, salinomycin-loaded polymer–lipid hybrid anti-HER2 nanoparticles.

Article Snippet: Human breast cancer cell lines BT-474 and MDA-MB-361 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: In Vivo, Injection

Journal: Molecular Cancer

Article Title: Phosphorylation of Ser 78 of Hsp27 correlated with HER-2/ neu status and lymph node positivity in breast cancer

doi: 10.1186/1476-4598-6-52

Figure Lengend Snippet: Effect of heregulin α1 (HRG α1) and p38 MAPK inhibitor (SB 203580) on Hsp27 phosphorylation . Cultures of cells from the BT474 breast cancer cell line were treated with HRG α1 for 10 and 30 min (A) or SB 203580 for 10 hours (B) and total cell lysates were extracted using M-PER reagent (Pierce). Equal amounts of proteins (20 μg) were separated by SDS-PAGE and transferred onto PVDF membrane. The membranes were blocked for 1 hour, followed by being probed with the respective primary antibodies (anti-Hsp27, anti-pSer 15 , anti-pSer 78 and anti-pSer 82 ), and HRP-conjugated secondary antibody. The signals were captured and their intensities were detected as described in Figure 2. The phosphorylation levels of pSer 15 , pSer 78 and pSer 82 were expressed as the ratios of intensity probed with phosphorylation site-specific antibody to the intensity probed with anti-Hsp27. Data with ± SD represents the average of triplicate experiments. C: control; SB: inhibitor SB203580. For the control of HRG-treated cells, untreated cells were cultured for 10 and 30 min and equal amounts of cellular proteins from both time intervals were mixed and used as control. For the control of inhibitor-treated cells, cells were treated with DMSO for 10 hours and cellular proteins were used as the control. * p < 0.05 (student t -test)

Article Snippet: The human BT474 breast cancer cell line was obtained from American Type Culture Collection, and maintained in modified Dulbecco's medium (HybriCare) supplemented with 10% fetal bovine serum (FBS) at 37°C in a humidified atmosphere of 5% CO 2 in an incubator.

Techniques: SDS Page, Cell Culture

Journal: Breast Cancer Research : BCR

Article Title: β1 integrin mediates an alternative survival pathway in breast cancer cells resistant to lapatinib

doi: 10.1186/bcr2936

Figure Lengend Snippet: Phosphorylated levels of β1 downstream kinases are increased upon acquisition of resistance to lapatinib (L) . ( A ) Parental (P) BT474 and ( B ) HCC1954 cells resistant to lapatinib (LRes), trastuzumab (TRes), and combination (LTRes) treatment strategies were developed by long-term exposure in 2D. Protein extracts were probed for β1, pHER2, pFAK, and pSrc, as well as totals.

Article Snippet: The human breast cancer cell line BT474 was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) and maintained as previously described [ ].

Techniques:

Journal: Breast Cancer Research : BCR

Article Title: β1 integrin mediates an alternative survival pathway in breast cancer cells resistant to lapatinib

doi: 10.1186/bcr2936

Figure Lengend Snippet: β1 blockade overcomes lapatinib resistance in BT474 cells in 3D culture . ( A ) Parental and LRes cells were plated on lrECM and treated ± lapatinib. ( B ) The β1 inhibitory antibody AIIB2--or IgG control--was applied to parental and LRes cells on Day 0 of plating on lrECM and allowed to grow for 12 days. ( C ) Parental and LRes cells were plated in lrECM with appropriate inhibitors, allowed to grow for five days, stained with Ki67 antigen or TUNEL labeling, and imaged. ( D ) 3D cultures of BT474 parental and LRes cells treated with AIIB2 or IgG were allowed to propagate for five days, then harvested for protein and immunoblotting. Densitometry measurements were normalized to total levels--except β1, which was normalized to β-actin--and are representative of three independent experiments. ( E ) The FAK inhibitor PF 573228 was applied to parental and LRes cells on Day 0 of plating on lrECM and cultures were allowed to grow for 12 days.

Article Snippet: The human breast cancer cell line BT474 was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) and maintained as previously described [ ].

Techniques: Staining, TUNEL Assay, Labeling, Western Blot

Journal: Breast Cancer Research : BCR

Article Title: β1 integrin mediates an alternative survival pathway in breast cancer cells resistant to lapatinib

doi: 10.1186/bcr2936

Figure Lengend Snippet: Percent growth inhibition of cells in response to HER-targeted therapies

Article Snippet: The human breast cancer cell line BT474 was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) and maintained as previously described [ ].

Techniques: Inhibition

Journal: Breast Cancer Research : BCR

Article Title: β1 integrin mediates an alternative survival pathway in breast cancer cells resistant to lapatinib

doi: 10.1186/bcr2936

Figure Lengend Snippet: HER2 and the β1 pathway play alternate roles in resistance to lapatinib-containing regimens, in comparison to trastuzumab . ( A ) Parental, LRes, and TRes cells were plated on lrECM in the presence of lapatinib and/or trastuzumab and assayed for response. ( B ) 3D cultures of parental, LRes, and TRes BT474 cells were harvested for protein and probed for phosphorylated and total HER receptors.

Article Snippet: The human breast cancer cell line BT474 was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) and maintained as previously described [ ].

Techniques:

Journal: Breast Cancer Research : BCR

Article Title: β1 integrin mediates an alternative survival pathway in breast cancer cells resistant to lapatinib

doi: 10.1186/bcr2936

Figure Lengend Snippet: β1 inhibition impedes colony grow of BT474 LRes and LTRes cells, but not parental or TRes . ( A ) Parental, LRes, LTRes, and TRes cells were plated in lrECM, subjected to HER2 and/or β1 inhibitors on Day 0, and propagated for 10 to 12 days. ( B ) The Hs_ITGB1_5 siRNA was validated both in 2D and 3D (top), transfected at 40 nMsi into parental, LRes, LTRes, and TRes cells, which were then grown on lrECM for 10 days (bottom).

Article Snippet: The human breast cancer cell line BT474 was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) and maintained as previously described [ ].

Techniques: Inhibition, Transfection

Journal: Oncoscience

Article Title: FS-93, an Hsp90 inhibitor, induces G2/M arrest and apoptosis via the degradation of client proteins in oncogene addicted and derived resistant cancer cells

doi:

Figure Lengend Snippet: A. Effects of FS-93 on cell proliferation. BT-474, NCI-H3122 and EBC-1 cells were treated with FS-93 at 25, 50 and 100nM or NVP-AUY922 at 100 nM for 72 h. Cell viability was assessed with SRB assay. Bars represent means±SD. B. Effects of FS-93 on degradation of client onco-proteins. BT-474, NCI-H3122 and EBC-1 cells were treated with FS-93 at 25, 50 and 100nM or NVP-AUY922 at 100 nM for 24 h. Cell lysates were prepared and analyzed with immunoblotting.

Article Snippet: Human breast cancer cell line BT-474 was obtained from American Type Culture Collection (Manassas, VA).

Techniques: Sulforhodamine B Assay, Western Blot

Journal: Oncoscience

Article Title: FS-93, an Hsp90 inhibitor, induces G2/M arrest and apoptosis via the degradation of client proteins in oncogene addicted and derived resistant cancer cells

doi:

Figure Lengend Snippet: A ., B. Effects of FS-93 on cell cycle distribution. BT-474, NCI-H3122 and EBC-1 cells were treated with FS-93 at 25, 50 and 100nM or NVP-AUY922 at 100 nM for 24 h. Cell cycle distribution was analyzed by FACS after propidium iodide staining. Representative images A. and quantification results B. were presented. Bars represent means±SD. C. Impacts of FS-93 on G2/M transition regulators. BT-474, NCI-H3122 and EBC-1 cells were treated with FS-93 at 25, 50 and 100nM or NVP-AUY922 at 100 nM for 24 h and cells lysates were immunoblotted with the indicated antibodies.

Article Snippet: Human breast cancer cell line BT-474 was obtained from American Type Culture Collection (Manassas, VA).

Techniques: Staining

Journal: Oncoscience

Article Title: FS-93, an Hsp90 inhibitor, induces G2/M arrest and apoptosis via the degradation of client proteins in oncogene addicted and derived resistant cancer cells

doi:

Figure Lengend Snippet: A. Effects of FS-93 on apoptosis induction. BT-474, NCI-H3122 and EBC-1 cells were treated with DMSO or FS-93 at 100 nM for 12, 24, 48 and 72h. Apoptosis was determined with annexin-V/PI assay. Bars represent means±SD. B. Impacts of FS-93 on apoptotic proteins. BT-474 cells were treated with DMSO or FS-93 at 100 nM for 12, 24, 48 and 72h and analyzed by immunoblotted with the indicated antibodies.

Article Snippet: Human breast cancer cell line BT-474 was obtained from American Type Culture Collection (Manassas, VA).

Techniques: