human breast cancer cell lines mda mb 231  (ATCC)

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: Ptch1 is expressed in breast cancer cell lines. A. PTCH1 mRNA expression (log 2 transformed) in various luminal and HER2 breast cancer cell lines (dark green) and in TNBC cell lines (other colors). TNBC cell lines are depicted according to the “Lehmann TNBC subtype” nomenclature : basal-like 1 (yellow), basal-like 2 (pale green), immunomodulatory (brown), luminal androgen receptor (dark pink), mesenchymal (pale pink) and mesenchymal stem-like (pink). B. PTCH1 protein expression in three TNBC cell lines. Western-blot was performed on 50 µg extracts from TNBC cell lines (MDA-MB-231, HCC-38 and MDA-MB-468) with antibodies directed against PTCH1. PTCH1 and GAPDH signals were quantified using ImageJ software. Data presented are the mean ± SEM of at least 3 independent experiments. Significance is calculated using Oneway ANOVA Turkey’s multiple comparisons test and attained at P < 0.05 (*).

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Expressing, Transformation Assay, Western Blot, Software

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: PTCH1 drug efflux inhibitor PAH increases the sensitivity of TNBC cells to chemotherapy . Cell viability was measured after 24 h or 48 h treatment with increasing concentration of docetaxel or doxorubicin respectively on MDA-MB-231, MDA-MB-468 and HCC-38 cell lines in the absence or the presence of 15µM PAH. IC 50 values (corresponding to the concentration of chemotherapy inducing 50% of cell death) were calculated. Data reported are the mean ± SEM of at least 3 independent experiments. Significance is attained at P < 0.05 (*).

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Concentration Assay

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: Other multidrug transporters are expressed in TNBC cell lines. A. P-gp and ABCG2 protein expression in three TNBC cell lines. Western-blot was performed on 50 µg extracts from each TNBC cell line (MDA-MB-231, HCC-38 and MDA-MB-468) with antibodies directed against P-gp or ABCG2 and GAPDH. B. P-gp, ABCG2 and GAPDH signals were quantified using ImageJ software. Data presented are the mean ± SEM of at least 3 independent experiments. Significance is calculated using Oneway ANOVA Turkey’s multiple comparisons test and attained at P < 0.05 (*). ** P < 0.01; *** P < 0.001; ns P > 0.05.

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Expressing, Western Blot, Software

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: PAH increases docetaxel efficacy against TNBC spheroids. MDA-MB-231 cells were plated in ultra-low attachment surface 24 well plates in complete medium and treated with increasing concentrations of docetaxel in the presence of DMSO (control, 0 PAH), PAH 10 µM or 30 µM. After 2 weeks pictures were taken using Cytation 5 cell imaging system from Biotek. The surface of spheroids was calculated and reported after normalization on the condition without docetaxel for each concentration of PAH.

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Control, Imaging, Concentration Assay

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: PAH addition to chemotherapy inhibits migration of TNBC cells. A. 100000 cells were seeded on membrane from Transwell plates. Cells were treated 24 h with doxorubicin ± 15µM PAH or 48 h with docetaxel ± 15µM PAH. After fixation and staining with crystal violet, wells were observed on a microscope with X5 objective. B. Migration was measured using a wound-healing assay. A wound was performed on MDA-MB-231 confluent cells seeded in 24 well plates. The medium was replaced by fresh medium containing 5 µM docetaxel in the absence of PAH, or the presence of 5 µM PAH. Two pictures were taken at two different points of each well immediately after wound, and 3 days after wound with 5X objective. The width of the wound was measured using ImageJ software and reported as final wound width/initial wound width in percentage. Data presented are the mean ± SEM of 3 independent experiments. Significance is attained at P < 0.05.

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Migration, Membrane, Staining, Microscopy, Wound Healing Assay, Software

Journal: Translational Oncology

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancers

doi: 10.1016/j.tranon.2026.102777

Figure Lengend Snippet: PTCH1 contributes to the efflux of doxorubicin from TNBC cells. A. PTCH1 protein expression (left panel) and intracellular doxorubicin (right panel) were analyzed 16 h after transfection of MDA-MB-231 cells with PTCH1-siRNA or negative-control-siRNA. PTCH1 and GAPDH western blot signals were quantified using ImageJ software (left panel). For doxorubicin accumulation measurements (right panel), MDA-MB-231 cells were grown on slides and transfected with PTCH1-siRNA or negative-control-siRNA. After incubation with 10 µM doxorubicin, 3 coverslips were fixed for doxorubicin loading control; the other coverslips were incubated with efflux buffer and fixed. B. Docetaxel inhibits the accumulation of doxorubicin in MDA-MB-231 cells. Cells on coverslip were incubated with 10 µM doxorubicin or 10 µM doxorubicin and 50 µM docetaxel. Doxorubicin fluorescence images were acquired by epifluorescence microscopy using a 40X objective, and doxorubicin fluorescence was quantified using ImageJ software for about 100 cells per condition per experiment. Histograms are the mean ± SEM of 3 independent experiments. Significance is attained at P < 0.05.

Article Snippet: Human breast cancer cell lines MDA-MB-231, MDA-MB-468 and HCC-38 were purchased from ATCC.

Techniques: Expressing, Transfection, Negative Control, Western Blot, Software, Incubation, Control, Fluorescence, Epifluorescence Microscopy

Journal: iScience

Article Title: CFIm25-dependent alternative polyadenylation in AKT2 mRNA programs macrophage polarization

doi: 10.1016/j.isci.2026.115791

Figure Lengend Snippet: CFIm25 regulates core macrophage functions in THP-1 cells (A) Phagocytic activity in CFIm25-overexpressing (OE) THP-1 cells. Flow cytometry analysis of fluorescently labeled IgG-coated latex bead uptake in M0, M1, and M2 polarized cells with or without CFIm25 OE. Data are shown as a percentage of fluorescent cells. (B) Phagocytic activity in M0, M1, and M2 polarized THP-1 cells with or without CFIm25 knockdown (KD). Data are shown as a percentage of fluorescent cells. (C) Chemotactic migration assay for CFIm25 OE cells. Quantification of cell migration through Trans -well membranes in response to LPS chemoattractant. Data represent the number of cells that have crossed the membrane, normalized to M0 control cells. (D) Chemotactic migration assay for CFIm25 KD cells. Data are normalized to M0 control cells. (E) Cytotoxic activity of CFIm25-overexpressing macrophages against cancer cells. Indirect co-culture assay assessing the viability of breast cancer (MDA-MB-231), lung cancer (A549), and pancreatic cancer (Mia-PaCa2) cells cultured in the lower chamber, with M1 or M2 polarized macrophages—with or without CFIm25 overexpression—seeded in the upper chamber separated by a permeable membrane. Data are presented as a percentage of viable cancer cells. (F) Cytotoxic activity of CFIm25-KD macrophages against cancer cells. Indirect co-culture assay with or without CFIm25 knockdown as described in E. Data for all panels are shown as mean ± SEM from three independent experiments, ∗∗ p < 0.01 and ∗ p < 0.05.

Article Snippet: MDA-MB-231; Breast Adenocarcinoma , ATCC , Cat# HTB-26.

Techniques: Activity Assay, Flow Cytometry, Labeling, Knockdown, Migration, Membrane, Control, Co-culture Assay, Cell Culture, Over Expression

Journal: bioRxiv

Article Title: ARHGEF7 S-glutathionylation promotes cancer cell migration through Rac1 activation

doi: 10.64898/2026.05.01.722049

Figure Lengend Snippet: ( a ) ARHGEF7 (human isoform-a, 646 residues, NM_001113513) domains, structure, and selected residues. The structure is an AlphaFold model (AF-Q14155-1-F1-model_v6_isoform_a) of ARHGEF7 showing the structured SH3, DH, and PH domains with cysteines and nearby residues. ( b ) Accessible surface areas (ASA) of cysteines in ARHGEF7. ( c ) Sequence alignments around C312 in ARHGEF7 orthologs. ( d ) Clickable glutathione approach. Cells expressing a glutathione synthetase mutant (GS M4) synthesize and produce clickable glutathione (N 3 -GSH) using endogenous γGlu-Cys and exogenous azido-Ala. Clickable glutathione forms S-glutathionylation with proteins, which are analyzed after click reactions. ( e - f ) Glutathionylation of ARHGEF7 WT and cysteine mutants in MDA-MB-231 cells in response to high glucose (HG, 25 mM) or low glucose (LG, 5 mM) conditions: cells were incubated for 20 h (n=3). ( g - h ) Glutathionylation of ARHGEF7 constructs in MDA-MB-231 cells upon adding EGF (n=3). EGF was incubated for 20 h. The lysates were used for click reactions with biotin-alkyne and analyzed by western blot with FLAG antibody before (input) and after (eluted) pull-down with streptavidin-agarose. Data represent the mean ± SD. The statistical difference was analyzed by one-way ANOVA and Tukey’s post-hoc test ( e - h ), where *p < 0.03, **p < 0.002, ***p < 0.0002, ****p < 0.0001.

Article Snippet: Breast cancer cell line MDA-MB-231 (ATCC, HTB-26TM) and breast cancer cell line MDA-MB-468 (ATCC, HTB-132TM) were maintained in Dulbecco’s Modified Eagle Medium with high glucose (DMEM, Cytiva, HycloneTM) supplemented with 10% Fetal Bovine Serum (FBS, Cytiva, HycloneTM) and 1% (100 U/mL) Penicillin-Streptomycin (Pen-strep, Thermo Scientific) in a humified incubator at 37°C in a 5% CO 2 environment.

Techniques: Sequencing, Expressing, Mutagenesis, Incubation, Construct, Western Blot

Journal: bioRxiv

Article Title: ARHGEF7 S-glutathionylation promotes cancer cell migration through Rac1 activation

doi: 10.64898/2026.05.01.722049

Figure Lengend Snippet: ( a - b ) In-vitro scratch migration analysis of MDA-MB-231 cells expressing ARHGEF7 construct in high glucose (HG, 25 mM) or low glucose (LG, 5 mM) conditions (n=4) ( a ) or in response to EGF (n=9) ( b ). Yellow indicates areas without cells. ( c - d ) Transwell invasion analysis of MDA-MB-231 cells expressing ARHGEF7 constructs in HL or LG (n=3) ( c ) or upon incubating EGF (n=3) ( d ). Cells were incubated in HG or LG ( a , c ) or with EGF for 20 h ( b , d ). A scale bar = 500 μm. Data represent the mean ± SD. The statistical difference was analyzed by two-way ANOVA and Tukey’s post-hoc test ( a - d ), where *p < 0.03, **p < 0.002, ***p < 0.0002, ****p < 0.0001.

Article Snippet: Breast cancer cell line MDA-MB-231 (ATCC, HTB-26TM) and breast cancer cell line MDA-MB-468 (ATCC, HTB-132TM) were maintained in Dulbecco’s Modified Eagle Medium with high glucose (DMEM, Cytiva, HycloneTM) supplemented with 10% Fetal Bovine Serum (FBS, Cytiva, HycloneTM) and 1% (100 U/mL) Penicillin-Streptomycin (Pen-strep, Thermo Scientific) in a humified incubator at 37°C in a 5% CO 2 environment.

Techniques: In Vitro, Migration, Expressing, Construct, Incubation

Journal: bioRxiv

Article Title: ARHGEF7 S-glutathionylation promotes cancer cell migration through Rac1 activation

doi: 10.64898/2026.05.01.722049

Figure Lengend Snippet: Analysis of ARHGEF7 protein interactions and Rac1-PAK1 downstream signaling. MDA-MB-231 cells expressing ARHGEF7 WT or C312S were incubated in high glucose (HG, 25 mM) or low glucose (LG, 5 mM) for 20 h. ( a ) ARHGEF7 S-glutathionylation increases its binding to Rac1. ARHGEF7 co-immunoprecipitation (co-IP) with Rac1 and PAK1 (n=3). ( b ) Rac1 is activated upon ARHGEF7 S-glutathionylation. Active Rac1 was enriched by purified GST-PBD (p21-binding domain derived from PAK1) and analyzed by western blots (n=3). ( c ) ARHGEF7 S-glutathionylation activates PAK1 and its downstream signaling. Phosphorylation levels of PAK1, LIMK1, and MEK1/2 were analyzed by western blot (n=3). ( d ) ARHGEF7 S-glutathionylation increases Rac1 localization at the cell periphery. The co-localization images of ARHGEF7 and Rac1 (left). Cells were fixed and analyzed by antibodies to FLAG (green) and Rac1/Cdc42 (red) (n=10 images). White arrows indicate the intense co-localization of ARHGEF7 and Rac1. Pearson’s correlation coefficients (right) were calculated after Intermodes thresholding to select for colocalization of ARHGEF7 and Rac1 at the cell periphery (see also Figure S6). A scale bar = 10 μm. Data represent the mean ± SD. The statistical difference was analyzed by one-way ANOVA and Tukey’s post-hoc test ( a-d ), where *p < 0.03, **p < 0.002, ***p < 0.0002, ****p < 0.0001.

Article Snippet: Breast cancer cell line MDA-MB-231 (ATCC, HTB-26TM) and breast cancer cell line MDA-MB-468 (ATCC, HTB-132TM) were maintained in Dulbecco’s Modified Eagle Medium with high glucose (DMEM, Cytiva, HycloneTM) supplemented with 10% Fetal Bovine Serum (FBS, Cytiva, HycloneTM) and 1% (100 U/mL) Penicillin-Streptomycin (Pen-strep, Thermo Scientific) in a humified incubator at 37°C in a 5% CO 2 environment.

Techniques: Expressing, Incubation, Binding Assay, Immunoprecipitation, Co-Immunoprecipitation Assay, Purification, Derivative Assay, Western Blot, Phospho-proteomics