过表达miR-299-3p抑制人肾癌细胞系增殖和迁移

目的探讨miR-299-3p在肾癌细胞系增殖、迁移和侵袭的作用及机制。方法RT-qPCR检测肾癌细胞系(786-O、769-P、ACHN和A498)和人正常肾小管上皮细胞系(HK-2)中miR-299-3p的表达;MTT法、细胞划痕实验、Transwell小室法检测miR-299-3p mimic对肾癌786-O细胞系增殖、迁移和侵袭的影响;在线生物信息学数据库对miR-299-3p的靶基因进行了预测。Western blot检测E-cadherin,N-cadherin、vimentin和MAP3K12的蛋白表达。结果miR-299-3p在肾癌细胞系中均低表达(P<0.05),且在786-O细胞中表达最低。过表达miR-299-3p可显著抑制肾癌细胞系786-O增殖、迁移、侵袭、体外成瘤和上皮间质转化(EMT),综合4个数据库的结果,发现预测的miR-299-3p的共同靶基为MAP3K12,荧光素酶报告基因验证了miR-299-3p与MAP3K12的靶向关系。miR-299-3p+MAP3K12共转染786-O细胞后,能够有效降低miR-299-3p mimic诱导的细胞迁移和侵袭的抑制作用。结论miR-299-3p在肾癌细胞中的作用机制可能是通过miR-299-3p靶向下调MAP3K12表达,调控EMT途径影响肾癌细胞增殖、迁移和侵袭。

Over-expression miR-299-3p inhibits proliferation and migration of renal carcinoma cell lines

Objective To explore the effect and mechanism of miR-299-3p effects on the proliferation,migration and invasion of renal cell carcinoma cell lines.Methods RT-qPCR was used to detect the expression of miR-299-3p in renal cancer cell lines(786-O,769-P,ACHN and A498)and human normal renal epithelial cell line(HK-2).MTT assay,wound healing assay,and Transwell assay were used to detect the effects of miR-299-3p on proliferation,migration,and invasion of 786-O cells.Online bioinformatics database was used to predict the target genes of miR-299-3p.Western blot analyzed the expression of E-cadherin,N-cadherin,vimentin and MAP3K12 in 786-O cells.Results miR-299-3p was lower expression in renal cancer cell lines and the lowest expression wasfound in 786-O cells.Over-expression of miR-299-3p significantly inhibited the proliferation,migration,invasion,tumor xenograft and EMT in 786-O cell and four databases predicted MAP3ZK12 as the common target of miR-299-3p.Dual luciferase reporter gene verified the targeting relationship between miR-299-3p and MAP3K12.miR-299-3p+MAP3K12 effectively reversed the inhibition of miR-299-3p mimic induced cell migration and invasion in 786-O cells.Conclusions The mechanism of miR-299-3p in renal cancer cells may be explained by down-regulated MAP3K12 to affect cell proliferation,migration,invasion and EMT pathway.