Effect Of MiR-519d-3p On Biological Function Of Hepatic Stellate Cells

Objective To investigate the effect of miR-519d-3p on the biological function of LX-2cell line.Methods 1.The molecular mimics(miR-519d-3p mimics),inhibitors(miR-519d-3p inbitor)and their negative controls(mimics NC and inhibitor NC)were transfected into LX-2cells,and the relative expression level of miR-519d-3p in LX-2 cells after tansfection was detected by real-time quantitative PCR;2.The use of real-time quantitative PCR and Western blot detection LX-2 after the transfection cells activation markers protein(alpha SMA)and alpha smooth muscle Ⅰ type Collagen,(CollagenⅠ)mRNA and protein expression level,analysis of miR-519d-3p’s influence on the LX-2 cell activation;3.CCK-8 kit was used to detect the effect of miR-519d-3p on LX-2 cell proliferation;4.Flow cytometry was used to detect the apoptosis rate of LX-2 cells after transfection,and the effect on the apoptosis of LX-2 cells was analyzed;5.cell scratch assay were used to detect the effect of miR-519d-3p on LX-2 cell migration.Results 1.After the transfection of miR-519d-3p into LX-2 cells,real-time quantitative PCR results showed that the relative expression of miR-519d-3p in the miR-519d-3p mimics group was significantly higher than that in the control group(con)and the mimics NC group(P<0.01).The relative expression level of miR-519d-3p in miR-519d-3p inhibitor group was significantly lower than that in control group and inhibitor NC group(P<0.01).2.miR-519d-3p transfection LX-2 cell,real time quantitative PCR and Western blot results show that the overexpression of miR-519d-3p inhibits the activation of LX-2 cells,compared with control group,the activation markers of alpha SMA and Collagen Ⅰ in mRNA and protein expression levels lower(P<0.01);By miR-519d-3p in LX-2 cells expressing quantity can promote the activation of LX-2 cells,compared with control group,the alpha SMA and Collagen Ⅰ in mRNA and protein expression level increased(P<0.01).3.After transfection of LX-2 cells with miR-519d-3p,overexpression of miR-519d-3p inhibited the proliferation of LX-2 cells compared with the control group(P<0.01);On the contrary,down-regulation of miR-519d-3p expression in LX-2 cells promoted the proliferation of LX-2 cells(P<0.01).4.After transfection with miR-519d-3p,the overexpression of miR-519d-3p significantly promoted the apoptosis of LX-2 cells compared with the control group(P<0.01);On the contrary,down-regulation of miR-519d-3p expression in LX-2 cells significantly inhibited apoptosis of LX-2 cells(P<0.01).5.After transfection of LX-2 cells with miR-519d-3p,cell scratch assay results showed that overexpression of miR-519d-3p inhibited the migration of LX-2 cells compared with the control group(P<0.01);On the contrary,down-regulation of miR-519d-3p expression in LX-2cells promoted the migration of LX-2 cells(P<0.01).Conclusion 1.miR-519d-3p can inhibit the activation and proliferation of LX-2 cells2.miR-519d-3p can promote the apoptosis of LX-2 cells3.miR-519d-3p can inhibit the migration of LX-2 cells...