Effect Of MiR-382 On Proliferation Of Rat Liver Cell Line BRL-3A

The liver is an organ with unique regenerative ability.Normally,it is in a resting state and rarely proliferates.When liver damage or partial hepatectomy(PH)occurs,hepatocytes are immediately induced and activated.The process of proliferation and liver regeneration restores its original volume,weight and function.In-depth study of the molecular mechanisms of liver regeneration has important guiding significance for revealing the pathogenesis of liver diseases and developing drugs for the treatment of liver diseases.microRNA is a single-stranded,non-coding,small RNAs with a hairpin structure that is highly conserved and has a hairpin structure,but has important cell-life activities such as cell proliferation,differentiation,apoptosis,and growth.It plays a regulatory role,complete or incomplete binding to the 3,UTR sequence of its target gene mRNA by base-pairing,weakening the post-transcriptional translation ability of mRNA or directly degrading the mRNA,thereby enabling the regulation of genes through post-transcriptional levels.Expression affects the physiological and biochemical activities of cells.Our previous studies showed that miR-382 is differentially expressed in rat regenerating liver and is a rat liver regeneration related miRNA.However,there have been no reports about the molecular mechanisms regulating the proliferation of rat hepatocytes.Therefore,this article mainly through the transfection of mir-382 mimic and inhibitor,overexpression and interference in the expression of miR-382,to detect its regulation of rat hepatocyte BRL-3A proliferation,in order to clarify the proliferation of mir-382 on regenerating hepatocytes The regulatory mechanism.MTT and EdU showed that miR-382 promoted the growth of BRL-3A cells after overexpression,miR-382 down-regulation inhibited the growth of BRL-3A cells;Flow cytometry showed that the number of cells in the S phase was significantly higher in the miR-382 overexpression group than in the NC group.The miR-382 interference group compared with the NC group,the number of cells in the S phase significantly reduced;the expression of PTEN-AKT signaling related genes/proteins was analyzed by mRNA and protein levels.RT-PCR and Western blot showed that PTEN was down-regulated after over-expression of miR-382,and P-AKT was up-regulated.The mRNA expression levels of Myc,Bcl2,and Ccnd1 were upregulated,and the apoptosis-related genes Caspase3 and Bax were detected.The level of mRNA expression was down-regulated,whereas the miR-382 interference group was contrary to the above results.Therefore,we speculate that miR-382 may promote the proliferation of rat hepatocytes BRL-3A by inhibiting the expression of PTEN and have the potential to promote liver regeneration and repair.