Regulation Of MiR-142-3p To Lactation Of Dairy Cow Mammary Epithelial Cells

Micro RNAs(mi RNAs) are endogenous, non-coding RNAs 21~25 nt in length which are highly conservative during evolution, and can make tatget m RNA clipped or transcription inhibited thereby translational repression or degradation of the transcript by pairing to sequences in the 3’ untranslated regions(3’-UTRs) of target m RNAs. mi RNAs have comprehensive and consequence Accommodation in mammals growth, development, cell differentiation, proliferation, apoptosis all that vital phenomena.and increasingly become hotspot and emphasis in various investigative territory.it is already known widely, mi R-142-3p can control cyclical variation of mice breast development lactation. but its accommodation has not reported in dairy cow mammary epithelial cells.In this study, we use mi R-142-3p as the study subject and dairy cow mammary gland as study model, to discover the effect of mi R-142-3p on mammogenesis and galactosis in dairy cow mammary epithelial cells. Aim to abundant theory and method about dairy cow galactosis function in contents. Therefore, possess important theory and practical significance. The experiment results showed that:①Real-time quantitative PCR results shows that mi R-142-3p expressed in mammary gland tissues of cows with different milk qualities. In the mammary gland tissues of cows producing high quality milk(H), the expression of mi R-142-3p was highest compared to pregnancy, and cows producing low quality milk(L). The results indicated that mi R-142-3p could regulate the cyclical changes of the dairy cow mammary gland development and lactation.②immunofluorescence technique results shows that prlr expressed in mammary gland tissues of cows with different milk qualities, in the mammary gland tissues of cows producing high quality milk(H) and pregnancy.③the function between prlr and mi R-142-3p was considered as a key point according the predict in targetscan(http://WWW.targetscan.org)prlr selected as target genes. detecting the expression of Prlr after treated with mi R-142-3p using q RT-PCR,when mi R-142-3p was overexpressed,Prlr was down-regulated,when mi R-142-3pwas inhibited,Prlr was up-regulated.④Using Western Blotting test the change expression of PRLR and related lactation albumen after transfection with mi R-142-3p mimics/inhibitor and mi R-142-3p mimics/inhibitor NC The expression of PRLR、STAT5、MAPK、AKT1、m TOR、SREBP1、PPARγ、Cyclin D1 and SCN2 were lowed after overexpress mi R-142-3p. However, When inhibition of mi R-142-3p, it is showed opposite results. Glut1 has no marked change.According to the results, we speculate that mi R-142-3p can regulate PRLR expression, which is associated with mammogenesis and galactosis signaling pathway molecule.⑤immunofluorescence technique results shows that after overexpress mi R-142-3p, PRLR location and expression in mammary gland tissues of cows has changed.EDU detected DNA multiplication capacity of DCMECs and flow cytometer detected mitotic cycle. Annexin V and PI double dyeing detect the apoptosis. The results showed that mi R-142-3p refrained cell viability and proliferation, and enhanced the apoptosis. However, When inhibition of mi R-142-3p, it is showed opposite results.⑥Because of the acceleration of bta-mi R-142-3p we found it could low triglyceride expression in culture solution, lactose has no obviously change;However, When inhibition of mi R-142-3p, it is showed opposite results,lactose has no obviously change. mi R-142-3p negative control breast component.In summary, we showed that mi R-142-3p is specifically involved in lactation of in dairy cow mammary epithelial cells by regulate target genes prlr,and down-regulates mammary gland development, cells reproductive activity,secretion of butterfat,the contents of lactation pathways proteins and has some influence on the Lactation function.