肾损伤分子-1特异性小干扰RNA对肾癌细胞株786—0细胞增殖及细胞周期的影响  

Effects of silencing kidney injury molecular-1 gene by small interfering RNA interference on prolif- eration and cell cycle of renal cell carcinoma 786-0 cells

作　　者：孟政雷 顾朝辉[1] 田凤艳[2] 贾占奎[1] 李冠儒[1] 孙科[1] 曾甫清[3] 杨锦建[1] 

机构地区：[1]郑州大学第一附属医院泌尿外科、河南省泌尿外科研究所、郑州市泌尿外科肿瘤分子生物学重点实验室,450052 [2]郑州大学第一附属医院儿科,450052 [3]华中科技大学同济医学院附属协和医院泌尿外科

出　　处：《中华实验外科杂志》2014年第6期1196-1198,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81100464、81200883）;中国博士后科学基金资助项目（2012M521410）;河南省卫生厅科技攻关项目（2011020039）

摘　　要：目的观察肾损伤分子-1（Kim-1）特异性小干扰RNA（siRNA）沉默Kim-1分子后，对786-0细胞增殖和细胞周期的影响。方法利用脂质体转染方法将对人Kim-1基因序列特异的siRNA（50nmol／L）转染进入786-0细胞中，转染后48h采用实时定量聚合酶链反应（Real-time PCR）检测Kim-1基因的表达，采用噻唑蓝（MTT）比色法检测细胞增殖，转染后48h采用流式细胞仪检测细胞周期。结果Kim-1-siRNA能有效下调786-0细胞中Kim-1基因的表达水平，抑制细胞生长，使细胞周期阻滞在G0／G1期[3组转染后96h的增殖抑制率分别为0、（3．80±1．24）％、（54．31±3．82）％，G0／G1期所占的比例分别为（38．44±1．82）％、（42．06±2．15）％、（63．69±2．87）％]，差异有统计学意义（P〈0．05）。结论siRNA能有效抑制786-0细胞中Kim-1基因的表达，Kim-1基因表达下调影响786-0细胞的增殖并将细胞周期阻滞于G0／G1期。Objective To investigate the effect of silencing kidney injury molecular-1 （ Kim-1 ） gene by small interfering RNA （siRNA） interference on proliferation and cell cycle of renal cell carcinoma 786-0 cells. Methods The siRNA （50 nmol/L） which targets human Kim-1 gene was transfected into 786-0 cells by lipofectamineTM 2000. After 48 h, the expression of Kim-1 was detected by quantitative realtime quantitative polymerase chain reaction （Real-time PCR） quantitatively. The proliferation ability of cells in each group was determined by methylthiazohetrazolium （MTT） assay respectively. After 48 h, the cell cycle was examined by flow cytometry. Results In 786-0 cells the siRNA which targets Kim-1 gene si- lenced the expression of Kim-1 significantly, inhibited the proliferation and induced cell cycle arrest at G0/ G1 phases [ for proliferation-inhibition rate 0, （3.80 ± 1.24 ）%, （54. 31± 3.82 ）% respectively; rate in G0/G1 phases （38.44 ± 1.82 ） %, （42. 06 ± 2. 15 ） %, （ 63.69± 2. 87 ） % respectively （ P 〈 0. 05 ） ]. Conclusion In 786-0 cells, Kim-1 expression can be inhibited significantly by siRNA, and the down-reg-ulation of Kim-1 expression could induce the proliferation inhibition and arrest of cell cycle at G0/G1 pha-ses.

关 键 词：肾细胞癌 肾损伤分子-1 小干扰RNA 增殖 细胞周期 

分 类 号：R737.11[医药卫生—肿瘤]