human normal mammary epithelial cells 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: Cancers

Article Title: Regulation of RORα Stability through PRMT5-Dependent Symmetric Dimethylation

doi: 10.3390/cancers16101914

Figure Lengend Snippet: Silencing PRMT5 decreases RORα and promotes EMT. ( a ) PRMT5 knockdown was confirmed by RT-PCR; ( b ) PRMT5 knockdown was confirmed by Western blotting; ( c ) SNAI mRNA levels were increased in PRMT5-silenced MCF10A cells. Results are presented as mean ± SEM; n = 3. * p < 0.05, independent Student’s t -test. ( d ) Phase contrast images of control and two PRMT5-knockdown MCF10A cell lines (shPRMT5-1 and shPRMT5-2) in 2D and 3D cultures. Bar: 50 µm; ( e ) EMT marker proteins were examined in control and PRMT5-silenced MCF10A cells.

Article Snippet: For the tissue culture plastic 2D cell culture, human normal mammary epithelial cell MCF10A cells (from American Type Culture Collection) were cultured in Dulbecco’s Modified Eagle’s Medium/F12 (DMEM/F12) (Sigma Aldrich, St. Louis, MO, USA) supplemented with 5% horse serum (Gibco, Brooklyn, USA), 0.5 mg/mL hydrocortisone (Sigma Aldrich), 20 ng/mL epidermal growth factor (EGF) (PEPROTECH, Rosemont, USA), 10μg/mL insulin (Sigma Aldrich), 100 ng/mL cholera toxin (Sigma Aldrich), and 1% penicillin–streptomycin (Sigma Aldrich).

Techniques: Knockdown, Reverse Transcription Polymerase Chain Reaction, Western Blot, Control, Marker

Journal: Cancers

Article Title: Regulation of RORα Stability through PRMT5-Dependent Symmetric Dimethylation

doi: 10.3390/cancers16101914

Figure Lengend Snippet: Silencing PRMT5 expression enhances cell invasion and migration. ( a ) Images of mammospheres in vector control or PRMT5-silenced MCF10A cells. Bar: 50 µm. ( b ) Mammosphere formation efficiency (MFE) was quantified in vector control or PRMT5-silenced MCF10A cells. Results are presented as mean ± SEM; n = 10; ** p < 0.01, determined by independent Student’s t test. ( c ) Transwell invasion analysis was conducted for both control and PRMT5-silenced MCF10A cells. Bar: 50 µm. ( d ) Invaded cells were quantified in vector control or PRMT5-silenced MCF10A cells. Results are presented as mean ± SEM; n = 3. ** p < 0.01, determined by independent Student’s t test. ( e ) Single-cell migration analysis was performed in control and PRMT5-silenced MCF10A cells. Results are presented as mean ± SEM; n = 100. ** p < 0.01, determined by independent Student’s t test.

Article Snippet: For the tissue culture plastic 2D cell culture, human normal mammary epithelial cell MCF10A cells (from American Type Culture Collection) were cultured in Dulbecco’s Modified Eagle’s Medium/F12 (DMEM/F12) (Sigma Aldrich, St. Louis, MO, USA) supplemented with 5% horse serum (Gibco, Brooklyn, USA), 0.5 mg/mL hydrocortisone (Sigma Aldrich), 20 ng/mL epidermal growth factor (EGF) (PEPROTECH, Rosemont, USA), 10μg/mL insulin (Sigma Aldrich), 100 ng/mL cholera toxin (Sigma Aldrich), and 1% penicillin–streptomycin (Sigma Aldrich).

Techniques: Expressing, Migration, Plasmid Preparation, Control

Journal: Cancers

Article Title: Regulation of RORα Stability through PRMT5-Dependent Symmetric Dimethylation

doi: 10.3390/cancers16101914

Figure Lengend Snippet: Elevated RORα protein levels in PRMT5-overexpressed cells suppresses clone growth and cell invasion. ( a ) PRMT5 overexpression was confirmed by RT-PCR; ( b ) PRMT5 overexpression was confirmed by Western blotting; ( c , d ) phase contrast images and quantification of control and PRMT5-overexpressed MCF10A cells in 3D culture. Bar: 100 µm. ( e , f ) Transwell invasion analysis of control and PRMT5-overexpressed MCF10A cells. Bar: 100 µm. Results are presented as mean ± SEM; n = 3. ** p < 0.01, independent Student’s t test.

Article Snippet: For the tissue culture plastic 2D cell culture, human normal mammary epithelial cell MCF10A cells (from American Type Culture Collection) were cultured in Dulbecco’s Modified Eagle’s Medium/F12 (DMEM/F12) (Sigma Aldrich, St. Louis, MO, USA) supplemented with 5% horse serum (Gibco, Brooklyn, USA), 0.5 mg/mL hydrocortisone (Sigma Aldrich), 20 ng/mL epidermal growth factor (EGF) (PEPROTECH, Rosemont, USA), 10μg/mL insulin (Sigma Aldrich), 100 ng/mL cholera toxin (Sigma Aldrich), and 1% penicillin–streptomycin (Sigma Aldrich).

Techniques: Over Expression, Reverse Transcription Polymerase Chain Reaction, Western Blot, Control

Journal: Experimental and Therapeutic Medicine

Article Title: Regulation of the stem‑like properties of estrogen receptor‑positive breast cancer cells through NR2E3/NR2C2 signaling

doi: 10.3892/etm.2023.12173

Figure Lengend Snippet: Expression of nr2e3 is upregulated in ER + breast adenoma tissue cells. Nr2e3 expression was analyzed in (A) ER + and (B) total breast adenoma tissues compared with normal tissue samples. Data were downloaded from The Cancer Genome Atlas database. (C) Western blotting showing the protein expression levels of NR2E3 in MCF10A and MCF7 cells, with GAPDH used as a loading control. BC, breast cancer; ER, estrogen receptor; NR2E3, nuclear receptor subfamily 2 group E member 3; FPKM, fragments per kilobase of exon model per million mapped fragments.

Article Snippet: Human normal mammary epithelial cells MCF10A (cat. no. CL-0525) and the ER + cell line MCF7 (cat. no. CL-0149) were purchased from Procell Life Science & Technology Co., Ltd.

Techniques: Expressing, Western Blot, Control

Journal: The journal of pathology. Clinical research

Article Title: Exosome-derived ANXA9 functions as an oncogene in breast cancer.

doi: 10.1002/cjp2.334

Figure Lengend Snippet: Figure 3. Validation of ANXA9 expression in BCA. (A) ANXA9 expression in adjacent non-tumor tissues and BCA tissues was detected by IHC staining. (B) RT-qPCR assay was to measure ANXA9 expression in adjacent non-tumor tissues and BCA tissues in 30 patients. (C) Western blot analysis of ANXA9 protein levels in adjacent non-tumor and tumor tissues of five patients. GAPDH was used as an internal control. (D) RT-qPCR and (E) western blot analysis of ANXA9 mRNA and ANXA9 protein levels in normal MCF10A cells, and BCA cell lines MCF7, SK-BR-3, and T-47D cells. Significance: **p < 0.01; ***p < 0.001.

Article Snippet: BCA cell lines MCF7, SK-BR-3, and T-47D cells and normal human mammary epithelial cell line MCF10A were purchased from ATCC (Manassas, VA, USA).

Techniques: Biomarker Discovery, Expressing, Immunohistochemistry, Quantitative RT-PCR, Western Blot, Control

Journal: The journal of pathology. Clinical research

Article Title: Exosome-derived ANXA9 functions as an oncogene in breast cancer.

doi: 10.1002/cjp2.334

Figure Lengend Snippet: Figure 5. ANXA9 is secreted by BCA tissue-derived exosomes. (A) Exosomes at different magnifications by transmission electron microscope. (B) Western blot was utilized to detect exosomal markers CD63, CD81, and CD9 protein levels in MCF10A, MCF, and T-47D cells cultured with or without exosomes. (C) ANXA9 mRNA expression level in MCF10A, MCF7, and T-47D cells cultured with or without exosomes was tested by RT-qPCR. (D, E) Proteinase and RNase protection assays were utilized to test ANXA9 mRNA and ANXA9 protein expression levels in MCF7 and T-47D cells cultured with exosomes and treated with/without RNase/Protease/Triton X-100. Significance: **p < 0.01; ***p < 0.001.

Article Snippet: BCA cell lines MCF7, SK-BR-3, and T-47D cells and normal human mammary epithelial cell line MCF10A were purchased from ATCC (Manassas, VA, USA).

Techniques: Derivative Assay, Transmission Assay, Microscopy, Western Blot, Cell Culture, Expressing, Quantitative RT-PCR

Journal: Hereditas

Article Title: Circular RNA circHMCU promotes breast tumorigenesis through miR-4458/PGK1 regulatory cascade

doi: 10.1186/s41065-023-00275-y

Figure Lengend Snippet: Expression and structure of circHMCU in BC. A Expression of circHMCU in Tumor tissues ( n = 66) and Normal tumor ( n = 66) was measured by qRT-PCR. B The expression of circHMCU in BC cell lines (MDA-MB-231 and MCF-7) and human normal mammary epithelial cell (MCF10A) were determined by qRT-PCR. C Schematic diagram showed the back-splicing was constituted with exon 3 and exon 4 in chr10: 74,474,868–74,475,660. D After RNase R treatment, the expression of circHMCU and linear HMCU were assessed by qRT-PCR. E After Actinomycin D treatment, the levels of circHMCU and linear HMCU were analyzed by qRT-PCR. * P < 0.05

Article Snippet: BC cell lines (MDA-MB-231 and MCF-7), human embryonic kidney cell line 293 T and human normal mammary epithelial cell line (MCF10A) were obtained from Procell (Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR

Journal: Hereditas

Article Title: Circular RNA circHMCU promotes breast tumorigenesis through miR-4458/PGK1 regulatory cascade

doi: 10.1186/s41065-023-00275-y

Figure Lengend Snippet: CircHMCU was a sponge of miR-4458. A The predicted binding site between circHMCU and miR-4458 was exhibited. B and C The binding relationship between circHMCU and miR-4458 was verified by dual-luciferase reporter assay and RIP assay. D The expression of miR-4458 in normal tissues ( n = 66) and tumor tissues ( n = 66) was tested by qRT-PCR. E miR-4458 expression was measured in BC cell lines (MDA-MB-231 and MCF-7) and human normal mammary epithelial cell (MCF10A). F Pearson analysis was conducted to analyze the correlation between circHMCU and miR-4458 in BC tissues. * P < 0.05

Article Snippet: BC cell lines (MDA-MB-231 and MCF-7), human embryonic kidney cell line 293 T and human normal mammary epithelial cell line (MCF10A) were obtained from Procell (Wuhan, China).

Techniques: Binding Assay, Luciferase, Reporter Assay, Expressing, Quantitative RT-PCR

Journal: Hereditas

Article Title: Circular RNA circHMCU promotes breast tumorigenesis through miR-4458/PGK1 regulatory cascade

doi: 10.1186/s41065-023-00275-y

Figure Lengend Snippet: MiR-4458 directly target PGK1. A The predicted binding site between miR-4458 and PGK1was exhibited. B The binding relationship between miR-4458 and PGK1 was verified by dual-luciferase reporter assay. C The expression of PGK1 in normal tissues ( n = 66) and tumor tissues ( n = 66) was tested by qRT-PCR. D The expression of PGK1 protein in in normal tissues and tumor tissues was tested by western blot. E PGK1 protein expression was measured in BC cell lines (MDA-MB-231 and MCF-7) and human normal mammary epithelial cell (MCF10A). F Pearson analysis was conducted to analyze the correlation between miR-4458 and PGK1 in BC tissues. G The expression of PGK1 protein was exhibited by western blot.* P < 0.05

Article Snippet: BC cell lines (MDA-MB-231 and MCF-7), human embryonic kidney cell line 293 T and human normal mammary epithelial cell line (MCF10A) were obtained from Procell (Wuhan, China).

Techniques: Binding Assay, Luciferase, Reporter Assay, Expressing, Quantitative RT-PCR, Western Blot