normal mammary epithelial cell line mcf10a  (ATCC)

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

Journal: Cancer Cell International

Article Title: β-Escin overcomes trastuzumab resistance in HER2-positive breast cancer by targeting cancer stem-like features

doi: 10.1186/s12935-022-02713-9

Figure Lengend Snippet: β-escin induces apoptosis in trastuzumab-sensitive and –resistant cells. A Chemical structure of β-escin. B The changes in morphology of BT474 and SKBR3 cells after treatment of β-escin (10-20 μM, 48 h) as observed by phase-contrast microscopy. C Representative phase-contrast images of JIMT-1 and MDA-MB-453 cells after treatment with β-escin (10-30 μM, 48 h). D , E Trastuzumab-sensitive SKBR3 and BT474 cells ( D ) and trastuzumab-resistant MDA-MB-453 and JIMT-1 cells ( E ) were treated with various concentrations of β-escin (1-100 μM) for 48 h, and cell viability was evaluated by MTS assay (**** p <0.0001). F , G BT474 ( F ) and JIMT-1 cells ( G ) were treated with β-escin (10-20 μM and 20-30 μM, respectively) for 48 h, and the percentages of cells in the sub-G1 phase were quantified using flow cytometry (** p <0.01). H , I The percentages of the early and late apoptotic cells in BT474 ( H ) and JIMT-1 cells ( I ) following exposure to β-escin (10-20 μM and 20-30 μM, respectively) were determined by annexin V/PI staining (**** p <0.0001). J Normal human mammary gland epithelial MCF10A cells were treated with β-escin (10-30 μM) for 48 h, and the sub-G1 fraction was analyzed (not significant, NS). The results are expressed as mean ± SEM after three independent experiments and analyzed by one-way ANOVA and Bonferroni’s post hoc test

Article Snippet: The human normal mammary epithelial MCF10A cell line (ATCC) was grown in MEGM supplemented with hEGF, hydrocortisone, insulin and bovine pituitary extract (SingleQuotsTM Kit, Lonza, CA) with 100 U/mL streptomycin-penicillin.

Techniques: Microscopy, MTS Assay, Flow Cytometry, Staining