breast adenocarcinoma cell line mda mb  (ATCC)

Journal: Cells

Article Title: IL-6 Blockade Enhances the Efficacy of CDK4/6 Inhibitor in BRCA1 -Mutant Triple-Negative Breast Cancer Cells

doi: 10.3390/cells14201602

Figure Lengend Snippet: Abemaciclib inhibited BRCA1 -mutant TNBC cell growth. SUM149, HCC1937, and MDA-MB-436 cells were treated with abemaciclib for 72 h. ( A ) Cell viability was determined by MTT assay. IC50 of abemaciclib was calculated and the IC50 ± SE values were listed at the bottom. ( B ) Cell proliferation was measured by BrdU cell proliferation assay. The drug concentrations used in the BrdU assay were selected based on the IC50 values determined in the MTT assay ( A ) to assess proliferation at biologically relevant doses. Data represents SD from three independent experiments (n = 3). ** p < 0.01, **** p < 0.0001.

Article Snippet: The human BRCA1 -mutant TNBC cell line MDA-MB-436 was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA; cat. no. HTB-13), SUM149 and HCC1937 were obtained from the University of Maryland Greenebaum Comprehensive Cancer Center Translational Laboratory (Baltimore, MD, USA).

Techniques: Mutagenesis, MTT Assay, BrdU Cell Proliferation Assay, BrdU Staining

Journal: Cells

Article Title: IL-6 Blockade Enhances the Efficacy of CDK4/6 Inhibitor in BRCA1 -Mutant Triple-Negative Breast Cancer Cells

doi: 10.3390/cells14201602

Figure Lengend Snippet: Bazedoxifene synergizes with abemaciclib in BRCA1 -mutant TNBC cells. Cell viability was measured by MTT assay in SUM149 ( A ), HCC1937 ( B ), and MDA-MB-436 ( C ) cells after treatment with bazedoxifene alone, abemaciclib alone, and the combination for 24 h (performed in triplicate assays). The CI values were reported in the bottom panel. *** p < 0.001, and **** p < 0.0001.

Article Snippet: The human BRCA1 -mutant TNBC cell line MDA-MB-436 was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA; cat. no. HTB-13), SUM149 and HCC1937 were obtained from the University of Maryland Greenebaum Comprehensive Cancer Center Translational Laboratory (Baltimore, MD, USA).

Techniques: Mutagenesis, MTT Assay

Journal: Cells

Article Title: IL-6 Blockade Enhances the Efficacy of CDK4/6 Inhibitor in BRCA1 -Mutant Triple-Negative Breast Cancer Cells

doi: 10.3390/cells14201602

Figure Lengend Snippet: GP130 knockdown enhances the sensitivity of abemaciclib in BRCA1 -mutant TNBC cells. ( A ) The knockdown efficiency of GP130 and its downstream P-STAT3 (Y705) and STAT3 were detected by western blot. GAPDH is used as a loading control. Western blot analysis was performed once due to sample limitations. ( B ) Cell viability of GP130 knockdown alone, abemaciclib alone, and the combination treatment was measured by MTT assay (performed in triplicate assays). * p < 0.05, ** p < 0.01, and *** p < 0.001. Original western blot images can be found in the .

Article Snippet: The human BRCA1 -mutant TNBC cell line MDA-MB-436 was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA; cat. no. HTB-13), SUM149 and HCC1937 were obtained from the University of Maryland Greenebaum Comprehensive Cancer Center Translational Laboratory (Baltimore, MD, USA).

Techniques: Knockdown, Mutagenesis, Western Blot, Control, MTT Assay

Journal: Cells

Article Title: IL-6 Blockade Enhances the Efficacy of CDK4/6 Inhibitor in BRCA1 -Mutant Triple-Negative Breast Cancer Cells

doi: 10.3390/cells14201602

Figure Lengend Snippet: Significant inhibitory effects of bazedoxifene and abemaciclib combination on the migration of BRCA1 -mutant TNBC cells. Wound healing assay was performed to evaluate cell migration in SUM149 ( A ), HCC1937 ( B ), and MDA-MB-436 ( C ) cells with bazedoxifene alone, abemaciclib alone, and the combination treatment for 24 h. Representative images of scratches with bazedoxifene alone (B15: 15 μM bazedoxifene; B20: 20 μM bazedoxifene), abemaciclib alone (A2.5: 2.5 μM abemaciclib), and the bazedoxifene and abemaciclib (B+A) combination treatment. The scale bar is 20 µM. The statistical analysis of cell invasion was shown in the right panel. Relative migration was quantified by measuring the wound area and statistical analysis was performed on data from triplicate assays. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The human BRCA1 -mutant TNBC cell line MDA-MB-436 was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA; cat. no. HTB-13), SUM149 and HCC1937 were obtained from the University of Maryland Greenebaum Comprehensive Cancer Center Translational Laboratory (Baltimore, MD, USA).

Techniques: Migration, Mutagenesis, Wound Healing Assay

Journal: Cells

Article Title: IL-6 Blockade Enhances the Efficacy of CDK4/6 Inhibitor in BRCA1 -Mutant Triple-Negative Breast Cancer Cells

doi: 10.3390/cells14201602

Figure Lengend Snippet: Significant inhibitory effects of bazedoxifene and abemaciclib combination on the invasion of BRCA1 -mutant TNBC cells. Transwell invasion assay was performed in triplicate to evaluate cell invasion. Representative images of invasive cells of SUM149 ( A ), HCC1937 ( B ), and MDA-MB-436 ( C ) with bazedoxifene alone, abemaciclib alone, and the combination treatment for 24 h (B15: 15 μM bazedoxifene; B20: 20 μM bazedoxifene; A1: 1 μM abemaciclib; A2.5: 2.5 μM abemaciclib and bazedoxifene and abemaciclib (B+A) combination treatment). The scale bar is 10 µM. The statistical analysis of cell invasion was shown in the right panel. * p < 0.05, and **** p < 0.0001.

Article Snippet: The human BRCA1 -mutant TNBC cell line MDA-MB-436 was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA; cat. no. HTB-13), SUM149 and HCC1937 were obtained from the University of Maryland Greenebaum Comprehensive Cancer Center Translational Laboratory (Baltimore, MD, USA).

Techniques: Mutagenesis, Transwell Invasion Assay

Journal: bioRxiv

Article Title: An inducible BRCA1 expression system with in vivo applicability uncovers activity of the combination of ATR and PARP inhibitors to overcome therapy resistance

doi: 10.1101/2025.08.05.668495

Figure Lengend Snippet: A ) Genetic alterations in the MDAMB436 cell line, as described in the Cancer Cell Line Encyclopaedia data update of 2019. B ) Western blot showing expression of RAD51B only in MDAMB436 cells stably transfected with a RAD51B expression construct. Vinculin was used as loading control. C ) Western blot showing expression of the tetracycline repressor (TetR) only in MDAMB436 or MDAMB436-B (+ RAD51B) cells transfected with the TetR construct (+). Vinculin was used as loading control. D ) Western blot showing expression of FLAG-tagged BRCA1 in the presence of tetracycline induction in MDAMB436-TR (+ TetR) or MDAMB436-B-TR (+ RAD51B + TetR) cells. GAPDH was used as loading control. E ) Dose-response curves of olaparib in colony formation assays in tetracycline-repressor (TR) expressing MDAMB436-B (+ RAD51B) cells with (+ BRCA1) or without (+ LacZ) BRCA1 complementation, cultured in the presence or absence of doxycycline (+/-dox). F ) Western blot showing immunoprecipitation of FLAG-tagged BRCA1 constructs (or LacZ control) stably transfected in MDAMB436-B-TR cells, in the presence or absence of doxycycline induction. FL: full length. GAPDH was used as input control.

Article Snippet: HEK-293T (CRL-3216) and MDAMB436 (HTB-130) cells were sourced from ATCC and grown in DMEM (Gibco) or RPMI 1640 (Sigma) respectively, both supplemented with 10% foetal bovine serum (FBS) (Sigma) and 1X GlutaMAX (Gibco).

Techniques: Western Blot, Expressing, Stable Transfection, Transfection, Construct, Control, Cell Culture, Immunoprecipitation

Journal: bioRxiv

Article Title: An inducible BRCA1 expression system with in vivo applicability uncovers activity of the combination of ATR and PARP inhibitors to overcome therapy resistance

doi: 10.1101/2025.08.05.668495

Figure Lengend Snippet: A ) Left panel: dose-response curves of olaparib in colony formation assays in tetracycline-repressor (TR) expressing MDAMB436 cells with or without RAD51B complementation (MDAMB436-B-TR and MDAMB436-TR, respectively) and with (+ BRCA1) or without (+ LacZ) BRCA1 complementation. Right panel: Logarithmic half-maximal inhibitory concentration (LogIC50) of olaparib for each cell line. B ) Left panel: dose-response curves of olaparib in colony formation assays in MDAMB436-B-TR cells expressing full-length BRCA1 (+FL), no BRCA1 (+LacZ) or different BRCA1 hypomorphs (+RING-less, +Δexon11, +BRCT-less). Right panel: Logarithmic half-maximal inhibitory concentration (LogIC50) of olaparib for each cell line. C ) Left panel: dose-response curves of olaparib in survival assays in MDAMB436-B-TR cells expressing full-length BRCA1 (+FL), no BRCA1 (+LacZ) or different BRCA1 hypomorphs (+C64R, +L1407P, +R1699Q). Right panel : Logarithmic half-maximal inhibitory concentration (LogIC50) of olaparib for each cell line. Data for BRCA1 hypomorphs with deletions in the RING (RING-less, M48 START) or BRCT (BRCT-less) domains are included for comparative purposes. D ) Left panel: dose-response curves of carboplatin in survival assays in MDAMB436-B-TR cells expressing full-length BRCA1 (+FL), no BRCA1 (+LacZ) or different BRCA1 hypomorphs (+RING-less, +Δexon11, +BRCT-less, +CC-mutant). Right panel : Logarithmic half-maximal inhibitory concentration (LogIC50) of carboplatin for each cell line. E ) Western blot of immunoprecipitation experiments in MDAMB436-B-TR cells expressing full length (+FL) or Δexon11 BRCA1 and exposed to different doxycycline doses. GAPDH was used as loading control. I = input; F = FLAG immunoprecipitation. F ) Logarithmic half-maximal inhibitory concentration (LogIC50) of olaparib at the different doses of doxycycline used. All data are from at least 3 biological replicates. Statistical analysis performed using One-Way ANOVA with Holm-Sidak multiple comparisons, * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001.

Article Snippet: HEK-293T (CRL-3216) and MDAMB436 (HTB-130) cells were sourced from ATCC and grown in DMEM (Gibco) or RPMI 1640 (Sigma) respectively, both supplemented with 10% foetal bovine serum (FBS) (Sigma) and 1X GlutaMAX (Gibco).

Techniques: Expressing, Concentration Assay, Mutagenesis, Western Blot, Immunoprecipitation, Control

Journal: bioRxiv

Article Title: An inducible BRCA1 expression system with in vivo applicability uncovers activity of the combination of ATR and PARP inhibitors to overcome therapy resistance

doi: 10.1101/2025.08.05.668495

Figure Lengend Snippet: A ) Tumour volume growth curves of MDAMB436 (BRCA1 mutant) cells or MDAMB436-B-TR cells expressing full length (+FL) BRCA1 in SCID mice. B ) Relative FLAG-BRCA1 mRNA expression in tumour samples from individual mice from the experiment in A. Expression is compared to samples taken from in vitro culture of the same cell lines and is normalized to that of GAPDH . C ) Dose-response efficacy of olaparib in the MDAMB436 (BRCA1m) xenograft model. D ) Dose-response efficacy of olaparib in the MDAMB436-B-TR + full length (FL) BRCA1 expressing xenograft model. E ) Dose-response efficacy of olaparib in the MDAMB436-B-TR + Δexon11 BRCA1 expressing xenograft model. Graphs depict geometrical mean (geomean) of tumour volume ± SEM and percentage TGI (tumour growth inhibition). Statistical significance was evaluated compared to the vehicle group using one-tailed t test (mice n = 8/group). Statistical significance is indicated as follows: * , P ≤ 0.05; ** , P ≤ 0.01; *** , P ≤ 0.001. F) Relative FLAG-BRCA1 mRNA expression in tumour samples from individual mice implanted with MDAMB436-BTR cells expressing the full length (FL) or Δexon11 BRCA1 constructs. Expression is normalized to that of GAPDH .

Article Snippet: HEK-293T (CRL-3216) and MDAMB436 (HTB-130) cells were sourced from ATCC and grown in DMEM (Gibco) or RPMI 1640 (Sigma) respectively, both supplemented with 10% foetal bovine serum (FBS) (Sigma) and 1X GlutaMAX (Gibco).

Techniques: Mutagenesis, Expressing, In Vitro, Inhibition, One-tailed Test, Construct

Journal: bioRxiv

Article Title: An inducible BRCA1 expression system with in vivo applicability uncovers activity of the combination of ATR and PARP inhibitors to overcome therapy resistance

doi: 10.1101/2025.08.05.668495

Figure Lengend Snippet: A ). Left panel: combination activity between olaparib and ceralasertib, SN-38, adavosertib or carboplatin in MDAMB436-B-TR cells expressing full length (+FL), Δexon11 or no BRCA1 (+LacZ) protein. Scores above 5 (marked by the dotted line on the graph) are described as synergistic based on the highest single agent (HSA) model. Right panel : representative examples of 6×6 combination matrices between olaparib and ceralasertib in MDAMB436-B-TR cells expressing full length (+FL) BRCA1 (top) or Δexon11 BRCA1 (bottom). In the Fits panels, values between 0-100 represent cytostatic effects. The excess (HSA) panels represent the values used to calculate synergy scores. Values coloured in pink/brown represent positive interactions. B ) Quantification of RAD51 foci formation in MDAMB436-B-TR cells expressing full length (+FL), Δexon11 or no BRCA1 (+LacZ) protein, irradiated (IR) or not with 5 Gy and treated or not with ceralasertib (0.3 µM for 24 hours prior to IR). Statistical analysis was performed using One-Way ANOVA with Sidak multiple comparisons, n=3 biological replicates with 2-3 technical replicates in each experiment; ns – not significant, * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001. C ) Quantification of γH2AX foci formation in MDAMB436-B-TR cells expressing full length (+FL), Δexon11 or no BRCA1 (+LacZ) protein, irradiated (IR) or not with 5 Gy and treated or not with ceralasertib (0.3 µM for 24 hours). Statistical analysis was performed using One-Way ANOVA with Sidak multiple comparisons, n=3 biological replicates with 2-3 technical replicates in each experiment, ns – not significant, * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001. D ) Efficacy of olaparib, ceralasertib or their combination in the MDAMB436-B-TR + Δexon11 BRCA1 expressing xenograft model. Graph depicts geometrical mean (geomean) of tumour volume ± SEM and percentage TGI (tumour growth inhibition). Statistical significance was evaluated compared to the vehicle group using one-tailed t test (mice n = 10/group). Statistical significance is indicated as follows: * , P ≤ 0.05; ** , P ≤ 0.01; *** , P ≤ 0.001.

Article Snippet: HEK-293T (CRL-3216) and MDAMB436 (HTB-130) cells were sourced from ATCC and grown in DMEM (Gibco) or RPMI 1640 (Sigma) respectively, both supplemented with 10% foetal bovine serum (FBS) (Sigma) and 1X GlutaMAX (Gibco).

Techniques: Activity Assay, Expressing, Irradiation, Inhibition, One-tailed Test