Journal: The Journal of Biological Chemistry

Article Title: Tumor-associated myoepithelial cells promote the invasive progression of ductal carcinoma in situ through activation of TGFβ signaling

doi: 10.1074/jbc.M117.775080

Figure Lengend Snippet: Tumor-associated myoepithelial cells activate the TGFβ/miR-10b-5p axis in DCIS cells and up-regulation of miR-10b-5p expression contributes to the coculture-enhanced EMT, invasiveness, and stemness of DCIS cells. A, PCR array profiling of miRNA expression in control and cocultured MCF10DCIS cells. miRNA expression data were plotted into a two-dimensional dot plot. Up-regulated (≥ 2-fold) and down-regulated (≤ −2-fold) miRNAs in sorted cocultured MCF10DCIS-GFP cells compared with the non-cocultured control are indicated. TGFβ-regulated miRNAs are depicted in the dot plot. B, qRT-PCR analysis of miR-10b-5p expression was performed on the sorted cell set as described in the legend to Fig. 4C. C, qRT-PCR analysis of miR-10b-5p expression was performed on RNA samples as described in the legend to Fig. 6D. D, qRT-PCR analysis of four EMT-programming genes was performed on MCF10DCIS cells stably overexpressing the control scramble, miR-10b, or miR-10b sponge RNA. Expression bar graph data shown in B–D were plotted based on triplicate experiments. E, inhibition of miR-10b by the sponge RNA partially suppresses coculture-promoted migratory and invasive activities of MCF10DCIS cells. Migration and invasion assays were performed on cocultured MCF10DCIS-GFP cells overexpressing the control scramble or miR-10b sponge RNA. Non-cocultured MCF10DCIS-GFP cells overexpressing the control scramble RNA served as a control. Non-cocultured and cocultured cells were sorted based on their GFP positivity before they were subjected to migration and invasion assays. Quantitative bar graph data (n = 3) were plotted as described in the legend to Fig. 1C. F, inhibition of miR-10b by the sponge RNA partially suppresses coculture-promoted CSC self-renewal of MCF10DCIS cells. Stem-cell sphere formation assays were performed on non-cocultured and cocultured cells as described in E. Quantitative CSC sphere formation data were plotted based on triplicate experiments. Error bars, S.D. *, p < 0.05; **, p < 0.01; ***, p < 0.001.

Article Snippet: miRNA expression profiling was carried out using the breast cancer miScript miRNA PCR arrays according to the manufacturer's instructions (Qiagen, Chatsworth, CA).

Techniques: Expressing, Control, Quantitative RT-PCR, Stable Transfection, RNA Expression, Inhibition, Migration

Journal: Cellular Oncology (Dordrecht)

Article Title: Identification of miR-10b, miR-26a, miR-146a and miR-153 as potential triple-negative breast cancer biomarkers

doi: 10.1007/s13402-015-0239-3

Figure Lengend Snippet: Expression of miRNAs in mammary cell lines. (a) miR-10b expression level, (b) miR-26a expression level, (c) miR-146a expression level and (d) miR-153 expression level. miRNA expression was determined by qRT-PCR in one benign mammary epithelium cell line (MCF10a) and nine tumor cell lines (two luminal cell lines: MCF7 and T47D and seven triple-negative cell lines: MDA-MB-231, SUM1315MO2, SUM1315-LXSN, SUM1315-BRCA1, MDA-MB-436, SUM149PT and HCC1937). Expression of miRNA was normalized using U6. A p -value < 0.05 is considered significant (between triple-negative and luminal cell lines)

Article Snippet: The Human Breast Cancer miScript miRNA PCR Array (MTHS-109ZE-4) was used according to the manufacturer’s protocol (Qiagen, France) for the profiling of 84 miRNAs known or predicted to alter in expression during breast cancer initiation and/or progression.

Techniques: Expressing, Quantitative RT-PCR

Journal: Cellular Oncology (Dordrecht)

Article Title: Identification of miR-10b, miR-26a, miR-146a and miR-153 as potential triple-negative breast cancer biomarkers

doi: 10.1007/s13402-015-0239-3

Figure Lengend Snippet: Profiling of miRNAs in human breast cancer cell lines by miScript miRNA PCR Array. Subgroups of cell lines are compared

Article Snippet: The Human Breast Cancer miScript miRNA PCR Array (MTHS-109ZE-4) was used according to the manufacturer’s protocol (Qiagen, France) for the profiling of 84 miRNAs known or predicted to alter in expression during breast cancer initiation and/or progression.

Techniques:

Journal: Cellular Oncology (Dordrecht)

Article Title: Identification of miR-10b, miR-26a, miR-146a and miR-153 as potential triple-negative breast cancer biomarkers

doi: 10.1007/s13402-015-0239-3

Figure Lengend Snippet: BRCA1 expression after miRNA mimic and miRNA inhibitor transfection in MDA-MB-231 and MCF7 cells. Expression of BRCA1 was determined by qRT-PCR in MDA-MB-231 and MCF7 cells transfected with Tmock (transfection reagent only), miR-146a, anti-miR-146a, miR-153, miR10-b and miR-26a. BRCA1 expression was normalized using 18S

Article Snippet: The Human Breast Cancer miScript miRNA PCR Array (MTHS-109ZE-4) was used according to the manufacturer’s protocol (Qiagen, France) for the profiling of 84 miRNAs known or predicted to alter in expression during breast cancer initiation and/or progression.

Techniques: Expressing, Transfection, Quantitative RT-PCR

Journal: Cellular Oncology (Dordrecht)

Article Title: Identification of miR-10b, miR-26a, miR-146a and miR-153 as potential triple-negative breast cancer biomarkers

doi: 10.1007/s13402-015-0239-3

Figure Lengend Snippet: Proliferation assay after siBRCA1, miRNA mimic and inhibitor transfection in MDA-MB-231 and MCF7 cells. Cells were transfected with Tmock (transfection reagent only), siBRCA1, miR-146a, anti-miR-146, miR-153, miR-10b and miR-26a. After 48 h, in vitro cell proliferation was evaluated using CCK-8. The absorbance was determined at 450 nm. All experiments were performed in triplicate

Article Snippet: The Human Breast Cancer miScript miRNA PCR Array (MTHS-109ZE-4) was used according to the manufacturer’s protocol (Qiagen, France) for the profiling of 84 miRNAs known or predicted to alter in expression during breast cancer initiation and/or progression.

Techniques: Proliferation Assay, Transfection, In Vitro, CCK-8 Assay

Journal: Cellular Oncology (Dordrecht)

Article Title: Identification of miR-10b, miR-26a, miR-146a and miR-153 as potential triple-negative breast cancer biomarkers

doi: 10.1007/s13402-015-0239-3

Figure Lengend Snippet: In silico expression analysis of miRNAs using TCGA data. Clinical and miRNA expression data for breast cancer were downloaded from The Cancer Genome Atlas (TCGA) database. The expression of four miRNAs (miR-10b, miR-26a, miR-146a and miR-153) was compared in 88 breast tumors with a negative ER, PR and HER2 status (i.e., triple negative phenotype) and in 431 breast tumors that were positive for at least one of the receptors. Student’s t- test was used to assess statistical differences in mean expression levels between these two groups

Article Snippet: The Human Breast Cancer miScript miRNA PCR Array (MTHS-109ZE-4) was used according to the manufacturer’s protocol (Qiagen, France) for the profiling of 84 miRNAs known or predicted to alter in expression during breast cancer initiation and/or progression.

Techniques: In Silico, Expressing