短发卡RNA靶向沉默TRPV2基因对EJ细胞增殖及迁移侵袭的影响

The effects of sh-RNA inhibiting TRPV2 gene expression on proliferation and invasion of hμman bladder cancer EJ cell line

目的:观察小干扰RNA(sh-RNA)抑制TRPV2基因的表达对人膀胱癌EJ细胞增殖及迁移侵袭的影响。方法:将人膀胱癌EJ细胞分为空白对照组、阴性对照组和sh-RNA干扰组进行实验。用lipofectamine2000介导针对TRPV2的sh-RNA转染EJ细胞。采用Western blot检测TRPV2蛋白表达的变化,MTT法检测细胞生长曲线,划痕法及transwell法检测细胞迁移侵袭能力。结果:sh-RNA TRPV2成功转染至EJ细胞。与空白对照组和阴性对照组相比较,sh-RNA TRPV2能诱导细胞体外增殖速度下降,并抑制细胞体外迁移能力(P<0.01)。结论:利用sh-RNA抑制TRPV2基因表达可有效抑制膀胱癌EJ细胞的体外增殖和迁移侵袭;TRPV2通道蛋白可能会成为膀胱癌治疗的新靶点。

Objective:To investigate the effects of sh-RNA inhibiting of the transient receptor potential vanilloid 2(TRPV2)channel protein on the invasion and proliferation in human urothelial carcinoma(UC)EJ cell line.Methods:EJ cells were divided into three groups:the blank control group,the negative control group,the sh-RNA-TRPV2 group.The vectors was transfected into EJ cells by lipofectamin2000.Western blotting was used to quantify TRPV2 protein levels.Then proliferation of EJ cells was determined by methyl thiazoltetrazolium(MTT)assay.The activities of motility and invasion of EJ cells were assessed by scratch assay and transwell chamber invasion assay in vitro.Results:In the sh-RNA-TRPV2 group,the TRPV2 protein levels were down regulated obviously.Compared to the blank control group and the negative control group,the proliferation abilities of the sh-RNA-TRPV2 group were inhibited,also the motility and invasion were inhibited in vitro(P<0.01).Conclusions:Sh-RNA-TRPV2 has negative effects on the proliferation and migration of EJ cells in vitro.TRPV2 may serve as an important target for the treatment of bladder neoplasms.