Knockdown of miRNA-376c-3p promotes cell growth in clear cell renal cell carcinoma
Objective To investigate the effect of miRNA-376c-3 on the growth of tumor cells in clear cell renal cell carcinoma(ccRCC).Methods Real-time fluorescence quantitative PCR(RT-PCR)was applied to detect the expression of miRNA-376c-3p in human ccRCC cell lines 786-O,Caki-1,SN12-PM6,ACHN,and the normal human proximal renal tubular epithelial cell line HKC.786-O cells were cultured in vitro.They were induced with blank control,transfection of miR-376c-3p inhibitor,transfection of miR-376c-3p mimics and transfection of negative control(NC).Expression level of miR-376c-3p was detected by RT-PCR.Cell proliferation was detected by immunofluorescence staining of Ki67 and colony formation.Cell apoptosis was detected by TUNEL staining.Positive expressions of apoptosis proteins Bax,Bcl2 and Bax/Bcl2 were detected by immunofluorescence.786-O cells induced with blank control,or transfected with miR-376c-3p inhibitor,miR-376c-3p mimics or NC were subcutaneously implanted into the right armpit of nude mice.Tumor volume and weight were calculated 3 weeks later.In vivo proliferation and apoptosis in nude mice were detected by immunofluorescence staining of Ki67 and TUENL staining,respectively.Results MiR-376c-3p was significantly downregulated in ccRCC cell lines 786-O,Caki-1,SN12-PM6 and ACHN compared with that of HKC cells(P＜0.05).Compared with those of controls,transfection of miR-376c-3p inhibitor significantly increased proliferative rate,colony formation rate,tumor volume and weight and positive expression rate of Ki67 in ccRCC xenografts of nude mice,but significantly decreased miR-376c-3p expression,Bax/Bcl-2,apoptotic rate and TUNEL-positive rate in ccRCC xenografts of nude mice(P＜0.05).Transfection of miR-376c-3p mimics significantly decreased proliferative rate,colony formation rate,tumor volume and weight and positive expression rate of Ki67 in ccRCC xenografts of nude mice,but significantly increased miR-376c-3p expression,Bax/Bcl-2,apoptotic rate and TUNEL-positive rate in ccRCC xenografts of nude mice(P＜0.05).The above indices were comparable between those induced with blank control and transfection of NC(P＞0.05).Conclusion knockdown of miRNA-376c-3p can weaken the proliferation and colony forming activity of ccRCC cells,promote their apoptosis,and inhibit the growth of transplanted tumors in nude mice.
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NSTL
万方数据
维普
