Effect of microRNA-214 on proliferation of mouse mesangial cells with lupus nephritis

YAO Feifei; SUN Liqiu; FANG Wei; WANG Huamin; YAO Dongsheng; CUI Rui; XU Jia; WANG Li; WANG Xiumei

doi:10.3969/j.issn.1002-266X.2016.48.005

Abstract：Objective To investigate the effect and molecular mechanism of microRNA-214 (miR-214) on the proliferation of mouse mesangial cells with lupus nephritis (LN).Methods The MRL-Faalpr/JNju mice (LN mice,n =20)and wild-type mice (n =20) were sacrificed and the glomerular mesangial cells were isolated and cultured.The relative expression of miR-214 was detected by real-time fluorescence quantitative PCR.The glomerular mesangial cells in the logarithmic phase were randomly divided into two groups,which were separately transfected by MiR-214 mimics (observation group) and NC mimics control (control group).The inhibitory rate of proliferation of mouse mesangial cells was detected by MTT.The target gene of miR-214 was predicted by bioinformatics analysis and verified by dual luciferase reporter assay.Furthermore,the mRNA and protein expression levels of YKL-40 were examined by real-time fluorescence quantitative PCR and Western blotting.LN mouse mesangial cells in the logarithmic phase were selected and then were transfected with siYKL-40 and negative control,respectively,and the inhibitory rate of cell proliferation was detected by MTT assay.Results The expression of miR-214 in the mouse mesangial cells of LN group was 0.35 ±0.09,which was significantly down-regulated as compared with that of the control group (1) (P ＜0.01).The expression of miR-214 in the miR-214 mimics group and NC mimics control group was 85.54 ± 12.07 and 1 (P ＜ 0.01).The inhibitory rate of proliferation in the observation group was higher than that of the control group on day 1,2 and 3 (P ＜0.05 or P ＜0.01).Compared with the control group,the relative expression of YKL-40 mRNA and protein decreased in the observation group (all P ＜ 0.05).Compared with that transfected with negative control,the proliferation inhibition rate of glomerular mesangial cells in the LN mice transfected with siYKL-40 increased (P ＜ 0.05 or P ＜ 0.01).Conclusions The expression of miR-214 is down-regulated in LN mouse mesangial cells.Up-regulation of miR-214 expression inhibits the proliferation of mouse mesangial cells,which may be mediated by YKL-40 down-regulation.