The Function And Regulation Targets Of MiR-142-3p In The Human Mammary Epithelial Cells Of MCF-10A

Micro RNAs are a series of non-coding and conservative RNAs, mi RNA contains approximately 23 nucleotides in length, which can regulate genes expression through complementary base pairing binding to target m RNA 3’ UTR(3’ untranslated region) in plants and animals, and then regulate genes expression at the post transcriptional level. most of the researches suggest that mi RNAs have an important influence on cell differentiation, cell growth, and cell death, Numerous studies have shown that mi R-142-3p have expressed in mammary gland,but it is not entirely clear that how regulation targets in human mammary epithelial cells.In this study, used mammary epithelial cells as experimental models,explore mi R-142-3p regulation targets and the role of in human mammary epithelial cells. RNA Binding Protein Immunoprecipitation(RIP)technology was used to screening the target gene of mi R-142-3p,At the same time,used the technology of EDU fluorescently labeled,flow cytometry,Transwell invasion assay and western blotting to detect how mi R-142-3p regulation cell differentiation, cell growth, and cell death in human mammary epithelial cells and the role of related signaling pathways.Results showed that:1)RIP experiment show that the target genes include fibroblast growth factor 5(FGF5)、metastasis associated in colon cancer 1(MACC1)、cyclin D1(CD1) take part in cell proliferation and differentiation. Validated Prolactin receptor(PRLR) as one of the target genes regulated of PRLR 3′UTR region, inhibited the expression of PRLR.2)After transfection of mi R-142-3p inhibitor and mi R-142-3p mimics in human mammary epithelial cells,PCR and Western blotting were used to detect gene expression and protein expression, the results shows that when mi R-142-3p was overexpressed, PRLR and lactation relative proteins were down-regulated, mi R-142-3p inhibition experiment showed opposite results.thus we can infer that mi R-142-3p can regulate the expression of PRLR, affected the function of mammary epithelial cells.3) mi R-142-3p was overexpressed in human mammary epithelial cells, used EDU and flow cytometer techniques to detect DNA proliferation and cell death, transwell plates were used to detect the ability of invasion and adhesion. through those different methods, we can verify that mi R-142-3p can inhibit the viability and multiplication capacity. mi R-142-3p was inhibited in human mammary epithelial cells,the results opposite.4) When mi R-142-3p was overexpressed theresults reveal that mi R-142-3p can down-regulate the contents of TG in the secretion, but there is little effect on lactose. when mi R-142-3p was inhibited,the contents of triglyceride in the secretion was up-regulated, still little effect on lactose secretion, the results confirm mi R-142-3p can regulate human mammary epithelial cells triglyceride secretion, but has no effective influence on lactose secretion.