小干扰RNA特异性沉默泛素特异肽酶22基因对膀胱癌细胞增殖的抑制作用  

The inhibitory effect of specific small interfering RNA against ubiquitin specific peptidase 22 gene on cell proliferation of bladder cancer cells

作　　者：吕磊[1] 曾甫清[1] 唐冬[1] 王智宇[1] 蒋国松[1] 肖行远[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院泌尿外科,武汉430022

出　　处：《肿瘤》2011年第1期17-21,共5页Tumor

基　　金：国家自然科学基金资助项目(编号:30972980)

摘　　要：目的:研究小RNA干扰泛素特异肽酶22(ubiquitin specific peptidase22,USP22)基因表达对膀胱癌EJ细胞增殖的影响。方法:设计并合成针对USP22基因的3条特异性小干扰RNA(small interfering RNA,siRNA)及阴性siRNA,并同时设立空白对照,然后用脂质体Translipid转染EJ细胞。通过实时荧光定量RT-PCR和Western印迹法分别检测转染前后EJ细胞中USP22mRNA和蛋白表达水平,MTT法和克隆形成实验检测细胞增殖率,FCM法检测细胞周期分布。结果:3条针对USP22基因的特异性siRNA均能抑制USP22mRNA和蛋白的表达,其中USP22-siRNA-1的沉默效率最高。转染USP22-siRNA-148h后,EJ细胞中USP22mRNA和蛋白表达水平分别下调65%和80%,而且细胞增殖明显受到抑制,细胞周期被阻滞于G1期。结论:USP22特异性siRNA能够有效沉默USP22基因表达,并显著抑制膀胱癌EJ细胞增殖。推测USP22基因可能成为治疗膀胱癌的一个新靶点。Objective： To investigate the effect of specific small interfering RNA （siRNA） against ubiquitin specific peptidase 22 （USP22） gene on the proliferation of bladder cancer cell line EJ. Methods： Three specific USP22 siRNAs and one negative siRNA were individually transfected into EJ cells for 48 h . The expression levels of USP22 mRNA and protein were analyzed by real-time fluorescence quantitative PCR （qRT-PCR） and Western blotting, respectively. The cell proliferation was determined by MTT assay and plate clone formation assay. The distribution of cell cycle was assessed by flow cytometry. Results： In comparison with the other two kinds of USP22 siRNAs, USP22-siRNA-1 was demonstrated to have the strongest silencing effect on USP22 gene expression in EJ cells at 48 h after transfection. The expression levels of USP22 mRNA and protein were reduced by 65% and 80%, respectively. The growth rate of EJ cells transfected with USP22-siRNA-1 was significantly decreased. Moreover, the cell cycle arrest in the G1 phase was observed when EJ cells were transfected with USP22-siRNA-1 for 48 h. Conclusion： Specific USP22 siRNAcan effectively inhibit the expression ofUSP22 gene and significantly suppress cell proliferation of bladder cancer EJ cells.

关 键 词：膀胱肿瘤 泛素巯基酯酶 RNA  小分子干扰 细胞增殖 细胞周期 泛素特异肽酶22 

分 类 号：R737.14[医药卫生—肿瘤]