siRNA抑制Cyclophilin A基因对喉鳞癌细胞Hep-2增殖和转移的影响  

Effects of silencing Cyclophilin A by siRNA on proliferation and metastasis in Hep-2 cells

作　　者：王致娟[1] 张跃东[1] 刘舟[1] 柏露[1] 周建荣[1] Wang Zhijuan Zhang Yuedong Liu Zhou Bai Lu Zhou Jianrong(Department of Otolaryngology, The First Affiliated Hospital of Chongqing Medical Universit)

机构地区：[1]重庆医科大学附属第一医院耳鼻喉科,重庆400016

出　　处：《重庆医科大学学报》2016年第10期1040-1045,共6页Journal of Chongqing Medical University

基　　金：重庆市卫生局医学科研重点资助项目(编号:2012-1-024)

摘　　要：目的:研究小干扰RNA(small interfering RNA,si RNA)抑制亲环素A(cyclopilin A,CypA)的表达对喉癌细胞株Hep-2增殖和侵袭迁移的影响及相关机制。方法:实验分为3组:化学合成的靶向CypA的si RNA转染喉癌细胞Hep-2(CypA si RNA组),转染阴性对照NC si RNA的细胞(NC si RNA组)和对照组(control组)。运用real-time PCR(RT-PCR)检测CypA及CD147m RNA的表达,Western blot检测CypA、CD147的蛋白表达。MTT法检测CypA对细胞生长曲线的影响;平板克隆形成实验检测CypA在喉癌细胞增殖中的作用;Transwell法检测CypA在喉癌细胞侵袭迁移中的作用。结果:CypA si RNA转染后的喉癌细胞株Hep-2中CypA、CD147的m RNA和蛋白表达水平均明显降低(P=0.000)。CypA si RNA转染后的Hep-2细胞生长缓慢,克隆形成被抑制,control组克隆数为(235.00±23.90)、NC si RNA组为(240.67±10.07)、CypA si RNA组为(129.33±14.22)(F=40.460,P=0.000);转染后Hep-2细胞的迁移(F=497.124,P=0.000)、侵袭(F=129.787,P=0.000)能力明显降低。结论:CypA表达下调后可抑制喉癌细胞株Hep-2的增殖和侵袭迁移能力,其机制可能与抑制CD147的表达有关。Objective：To investigate the effect of silencing cyclopilin A（CypA）expression on proliferation,migration and invasion of laryngeal carcinoma cell line Hep-2 and its mechanism. Methods：si RNAs targeting CypA and negative control si RNA were transfected into Hep-2 cells. Real-time PCR（q RT-PCR）was used to detect the m RNA expression of CypA and CD147. Protein expression of CypA and CD147 was detected by Western blot. The impact of CypA on cell growth was detected by MTT assay. Colony formation assay was used to examine the role of CypA in cell proliferation. Transwell test was used to test the invasion and migration ability of Hep-2 cells.Results：si RNA CypA markedly decreased both m RNA and protein expression of CypA and CD147（P=0.000）. Knockdown of CypA significantly down-regulated the m RNA and protein expression of CD147（P=0.000）. si RNA CypA reduced the proliferation and colony formation,and the number of colonies was significantly lower in CypA si RNA group（129.33±14.22）than in control group（235.00±23.90）and NC si RNA group（240.67±10.07）（F=40.460,P=0.000）;the migration（F=497.124,P=0.000）and invasion（F=129.787,P=0.000）ability of Hep-2 cells was distinctly decreased. Conclusion：Down-regulation of CypA can inhibit the proliferation,migration and invasion ability of Hep-2 cells,which may due to the decrease of CD147.

关 键 词：小干扰RNA 亲环素A HEP-2细胞株 增殖 侵袭 

分 类 号：R739.65[医药卫生—肿瘤]