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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。
AIM: To investigate the effect of small interfering RNA (Ubiquitin specific peptidase 22, USP22) gene on the proliferation of bladder cancer EJ cells. Methods: Three small interfering RNAs (siRNAs) against the USP22 gene and negative siRNAs were designed and synthesized. At the same time, a blank control was set up and transfected into EJ cells with liposome Translipid. The expression of USP22 mRNA and protein in EJ cells before and after transfection were detected by real-time fluorescence quantitative RT-PCR and Western blot respectively. The cell proliferation rate was detected by MTT assay and colony formation assay. The cell cycle distribution was detected by FCM. Results: Three specific siRNAs targeting USP22 gene both inhibited the expression of USP22 mRNA and protein. Among them, USP22-siRNA-1 had the highest silencing efficiency. After transfected with USP22-siRNA-148h, the expression levels of USP22 mRNA and protein in EJ cells were down-regulated by 65% and 80%, respectively, and cell proliferation was significantly inhibited and the cell cycle was arrested in G1 phase. Conclusion: USP22 specific siRNA can effectively silence USP22 gene expression and significantly inhibit bladder cancer EJ cell proliferation. Speculated that the USP22 gene may be a new target for the treatment of bladder cancer.