The methylation level of miR-4756 in prostate cancer tissue and its effect on the proliferation and migration of prostate cancer cells
Objective To analyze the methylation level of miR-4756 in prostate cancer tissues,and to explore the effect and functional mechanism of miR-4756 on the proliferation and migration of prostate cancer cells.Methods The iMETHYL database was used to analyze the methylation condition of mi R-4756 in prostate cancer tissues.Real-time fluorescent quantitative PCR(qRT-PCR)was used to detect the expression of miR-4756 in prostate cancer 22Rv1,DU-145,PC-3,and C4-2B cells.The 5-aza-2'-deoxycytidine(5-Aza-CdR)reagent was used to treat prostate cancer cells.The effect of 5-Aza-CdR on the expression of miR-4756 in prostate cancer cells was detected by qRT-PCR.22Rv1 cells were transfected with miR-4756 mimic to upregulate the expression of miR-4756.Colony formation assay and scratch healing assay were used to detect the effect of the up-regulated miR-4756 on the proliferation and migration of 22Rv1 cells.The dual-luciferase reporter assay was used to verify the targeting relationship between miR-4756 and glypican-6(GPC6).qRT-PCR and Western blot were used to detect the effect of the up-regulated miR-4756 on the expression of GPC6 gene in 22Rv1 cells.Western blot was used to detect the effect of the up-regulating miR-4756 on the expression of Wnt signaling pathway proteins Wnt1,Wnt2,ROR1 and ROR2 in 22Rv1 cells.Results The methylation level of miR-4756 in prostate cancer tissues was significantly higher than that in adjacent tissues(P＜0.01).Compared with human normal prostate epithelial cells(RWPE-1),the miR-4756 expression was significantly decreased in prostate cancer cells(P＜0.01).5-Aza-CdR reagent could promote the expression of miR-4756 in prostate cancer cells(P＜0.01).The number of 22Rv1 cells clones formed in the miR-4756 group was lower than that in the control group.The migration rates of 22Rv1 cells in the miR-4756 group were lower than those in the control group(P＜0.01).mi R-4756 can target GPC6 messenger RNA(mRNA)(P＜0.01).Compared with the control group,the expression of GPC6 protein in 22Rv1 cells was significantly inhibited by the upregulated miR-4756,and the expressions of Wnt signaling pathway proteins Wnt1,Wnt2,ROR1,and ROR2 were significantly reduced(P＜0.01).Conclusions The methylation level of miR-4756 in prostate cancer tissue is higher than that in adjacent tissue,and up-regulation of miR-4756 inhibits the proliferation and migration of prostate cancer cells by targeted down-regulation of GPC6 expression in prostate cancer 22Rv1 cells.
万方数据
维普
