siRNA沉默USP22基因对胃癌细胞增殖的抑制作用  

SiRNA-mediated silencing of the USP22 gene inhibits cell proliferation in human gastric cancer cell line AGS

作　　者：邓美洲[1] 陶凯雄[1] 王国斌[1] 刘兴华[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院腔镜外科,湖北省武汉市430022

出　　处：《世界华人消化杂志》2011年第19期1985-1989,共5页World Chinese Journal of Digestology

摘　　要：目的:探讨小干扰RNA(siRNA)沉默泛素特异性肽酶22(ubiquitin specific peptidase22,USP22)对胃癌细胞增殖的影响.方法:针对USP22基因设计3条siRNA及阴性siRNA,用脂质体Lipofectamine 2000转染胃癌AGS细胞,通过实时定量PCR和Western blot检测转染后AGS细胞USP22基因中mRNA和蛋白表达水平的变化情况,流式细胞术检测细胞周期分布变化情况,CCK8法检测细胞增殖率及抑制率.结果:转染48h后,3条siRNA均能显著抑制USP22 mRNA和蛋白的表达.其中,以转染USP22 siRNA3后效果最明显,mRNA和蛋白表达分别下降80.47%±2.99%和79.40%±3.58%.细胞增殖明显受到抑制,USP22 siR-NA3组细胞增殖抑制率为27.33%±3.49%.细胞周期中G0/G1期细胞增多,S期细胞减少.结论:采用RNA干扰技术能够有效地沉默USP22基因的表达,并显著抑制胃癌细胞的增殖.AIM：To evaluate the impact of silencing of the USP22 gene by small interfering RNA（siRNA） on the proliferation of human gastric cancer AGS cells. METHODS：Three USP22-specific siRNAs and a negative siRNA were designed and transfected into AGS cells using Lipofectamine 2000.Quantitative real-time PCR（qRT-PCR）and Western blot were utilized to detect the expression levels of USP22 mRNA and protein,respectively.Cell proliferation was measured using Cell Counting Kit-8（CCK-8）.The distribution of cell cycle was determined by flow cytometry. RESULTS：All three USP22-specific siRNAs could silence the expression of the USP22 gene.Forty-eight hours after transfection,the expression levels of USP22 mRNA and protein were reduced by 80.47%±2.99%and 79.40%±3.58%, respectively;the reduced rate of cell proliferation was 27.33%±3.49%;and the proportion of gastric cancer cells arrested in G0/G1 phase increased significantly,while those arrested in S phase decreased significantly. CONCLUSION：Transfection of USP22-specific siRNAs could effectively inhibit the expression of the USP22 gene and significantly suppress cell growth in human gastric cancer cell line AGS.

关 键 词：泛素特异性肽酶22 小干扰RNA 胃癌 AGS细胞 细胞增殖 

分 类 号：R735.2[医药卫生—肿瘤]