human mammary epithelial cell line mcf10a  (ATCC)

Journal: Frontiers in Pharmacology

Article Title: LITAF suppresses breast cancer and paclitaxel resistance by ubiquitinating and degrading PCMT1 to inhibit COX-2-dependent arachidonic acid metabolism

doi: 10.3389/fphar.2026.1706420

Figure Lengend Snippet: PCMT1 is involved in PTX resistance of BC cells, a process potentially involving COX-2-mediated AA metabolism. (A) RT-qPCR analysis of PCMT1 mRNA levels in tumor tissues and paired adjacent normal tissues from 30 BC patients treated with PTX. (B) Immunoblotting analysis of PCMT1 and COX-2 protein levels in tumor tissues and paired adjacent normal tissues from PTX-treated BC patients (n = 10; 5 PTX-sensitive and 5 PTX-resistant). (C) Correlation between PCMT1 and COX-2 expression was assessed using the Pearson correlation coefficient. (D) RT-qPCR analysis of PCMT1 mRNA levels in MCF10A, BC cells (MDA-MB-231, MCF-7), and PTX-resistant cells (MDA-MB-231/PTX, MCF-7/PTX) (n = 3). (E) Immunoblotting analysis of PCMT1 and COX-2 protein levels in BC cells and PTX-resistant cells (n = 3). (F) IC50 values of BC cells and PTX-resistant cells assessed using the CCK-8 assay (n = 3). Values are represented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Human normal mammary epithelial cells MCF10A (ATCC, USA) were cultured in DMEM/F12 (Procell) supplemented with HS (5%), EGF (20 ng/mL), hydrocortisone (0.5 μg/mL), insulin (0.5 μg/mL), NEAA (1%), and P/S (1%).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, CCK-8 Assay

Journal: Journal of Medicinal Chemistry

Article Title: Exploring Gallium(III) Complexes as Emerging Therapeutic Candidates for Breast Cancer

doi: 10.1021/acs.jmedchem.5c03480

Figure Lengend Snippet: (A) Dose–response curves of Ga3 and Ga6 compared to cisplatin in nontransformed cell lines HaCaT and MCF10A. (B) Selectivity Index values for Ga3 , Ga6 and cisplatin. Data represent the mean ± SD from three independent experiments performed in triplicate.

Article Snippet: The cell lines MCF7, SKBR3, BT549, and MDA-MB-231, the immortalized nontransformed keratinocyte cell line HaCaT, and the human mammary epithelial cell line MCF10A were acquired from ATCC.

Techniques:

Journal: Journal of Medicinal Chemistry

Article Title: Exploring Gallium(III) Complexes as Emerging Therapeutic Candidates for Breast Cancer

doi: 10.1021/acs.jmedchem.5c03480

Figure Lengend Snippet: Matrigel assay. (A) 3D structures of MCF7, MDA-MB-231 and MCF10A seeded in Matrigel matrix and treated with compound at indicated doses for 72-h. Scale bar = 250 μm. (B) The diameter of 3D structures was quantified using ImageJ. Data are the average ± SEM of three independent experiments. The values for the statistical analyses are * p ≤ 0.05; *** p ≤ 0.001.

Article Snippet: The cell lines MCF7, SKBR3, BT549, and MDA-MB-231, the immortalized nontransformed keratinocyte cell line HaCaT, and the human mammary epithelial cell line MCF10A were acquired from ATCC.

Techniques: Matrigel Assay

Journal: Breast Cancer Research : BCR

Article Title: Deubiquitination and stabilization of EIF4A3 by OTUB2 contributes to TPI1-mediated glycolysis and TNBC progression

doi: 10.1186/s13058-026-02260-5

Figure Lengend Snippet: Elevated OTUB2 expression in TNBC tissues and cell lines. A The Cancer Genome Atlas (TCGA) database was used to analyze OTUB2 expression in breast cancer tissues and TNBC tissues. * p < 0.05; *** p < 0.001. B and C qPCR and western blot detected the expression of OTUB2 in TNBC tissues and para-carcinoma tissues ( n = 30 samples per group). ** p < 0.01;*** p < 0.001. D and E qPCR and western blot detected the expression of OTUB2 in MCF10A, MDA-MB-231, MDA-MB-453, MDA-MB-468, SUM-159, and BT-549 cells ( n = 3 per group). * p < 0.05; ** p < 0.01; *** p < 0.001. Results are expressed as mean ± SD. Statistical analyses were analyzed using a Student’s t-test in A (left), B, and C, while one-way ANOVA followed by Tukey’s multiple-comparisons test were used to analyze differences between groups in A (right), D, and E

Article Snippet: MCF10A mammary epithelial cells were purchased from ATCC.

Techniques: Expressing, Western Blot

Journal: Breast Cancer Research : BCR

Article Title: Deubiquitination and stabilization of EIF4A3 by OTUB2 contributes to TPI1-mediated glycolysis and TNBC progression

doi: 10.1186/s13058-026-02260-5

Figure Lengend Snippet: OTUB2 stabilizes EIF4A3 by deubiquitination. A TCGA database was used to analyze EIF4A3 expression in breast cancer tissues and TNBC tissues. *** p < 0.001. B Western blot detected the expression of EIF4A3 in TNBC tissues and para-carcinoma tissues ( n = 30 samples per group). ** p < 0.01. C Western blot detected the EIF4A3 protein in MCF10A, MDA-MB-231, MDA-MB-453, MDA-MB-468, SUM-159, and BT-549 cells. * p < 0.05; ** p < 0.01. D and E MDA-MB-468 cells were infected with sh-OTUB2, sh-NC, oe-OTUB2, and oe-NC, respectively. Relative mRNA and protein expressions of OTUB2 and EIF4A3 were detected. ** p < 0.01; *** p < 0.001. F Co-IP assay for measurement of interaction between OTUB2 and EIF4A3. OTUB2 and EIF4A3 protein expressions were evaluated in anti-OTUB2 complex or anti-IgG by western blot. G MDA-MB-468 cells infection with sh-OTUB2 or sh-NC were treated with CHX for 0, 2, 4, 6, 8 h, respectively. EIF4A3 protein was measured by western blot. * p < 0.05; ** p < 0.01. H MDA-MB-468 cells infection with sh-OTUB2 or oe-OTUB2 were treated with MG132 for 6 h. Ubiquitination assay was used to test the effect of OTUB2 on EIF4A3 ubiquitination. I In vitro ubiquitination assay. Purified V5-ubiquitin and Flag-EIF4A3 were incubated with either HA-OTUB2 or HA-OTUB2 C51S . Then, Flag-U2AF2 proteins were further purified with anti-Flag antibody and immunoblotted with antibodies against V5 and Flag. n = 3 per group; Results are expressed as mean ± SD. Statistical analyses were analyzed using a Student’s t-test in A (left) and B, while one-way ANOVA followed by Tukey’s multiple-comparisons test were used to analyze differences between groups in A(right), C-E, and G

Article Snippet: MCF10A mammary epithelial cells were purchased from ATCC.

Techniques: Expressing, Western Blot, Infection, Co-Immunoprecipitation Assay, Ubiquitin Proteomics, In Vitro, Purification, Incubation