人乳头瘤病毒16E6基因沉默对鼻咽癌细胞侵袭转移能力及细胞周期的影响  

Human papillomavirus 16 E6 gene silencing suppresses invasion and cell cycle of nasopharyngeal carcinoma cells in vitro

作　　者：沈娜[1] 张潜英[1] 余晓燕[1] 叶琳[1] 陈鸿雁[1] 

机构地区：[1]重庆医科大学附属第一医院耳鼻咽喉科,重庆400016

出　　处：《第三军医大学学报》2009年第18期1757-1760,共4页Journal of Third Military Medical University

基　　金：重庆市卫生局资助项目(07-1-021)~~

摘　　要：目的观察针对人乳头瘤病毒(human papillomavirus,HPV)16E6基因的RNA干扰(RNA interfering,RNAi)对人鼻咽癌(nasopharyngeal carcinoma,NPC)HNE-1细胞侵袭转移能力及细胞周期的影响,探讨HPV16E6基因作为鼻咽癌基因治疗标靶的可行性。方法根据基因库上的HPV16E6mRNA序列,设计合成针对HPV16E6的siRNA,转染HNE-1细胞,采用RT-PCR及Westernblot分析E6基因的表达,利用侵袭实验检测细胞的侵袭能力,MTT法检测细胞的生长增殖,流式细胞术检测细胞周期。结果针对HPV16E6的siRNA下调鼻咽癌HNE-1细胞株E6mRNA和蛋白表达水平,分别下调66.3%、56.7%(P<0.05);侵袭实验结果显示,siRNA转染组的穿膜细胞数为(45.3±4.0),显著低于阴性对照组[(147.7±4.7),P<0.05];转染组细胞的增殖受到抑制,与阴性对照组相比差异有统计学意义(P<0.05);流式细胞术显示细胞阻滞于G0/G1期。结论siRNA可以有效干扰鼻咽癌HNE-1细胞内HPV16E6基因的表达,进而影响HNE-1细胞的侵袭转移能力,并抑制肿瘤细胞的生长增殖,使细胞周期重新分布。Objective To observe the effect of HPV16 E6-targeted RNA interference （RNAi） on the invasion, metastasis ability and cell cycle of human nasopharyngeal carcinoma （NPC） cell line HNE-1 and investigate the feasibility of using HPV16 E6 gene as therapeutic target for treatment of NPC. Methods According to HPV16 mRNA sequence in the Genbank, the siRNA against HPV16 E6 were designed, synthesized and transfected into HNE-1 cells. The mRNA and protein expression of HPV16 E6 were confirmed by RT-PCR and Western blotting, respectively. The invasion ability was determined by Transwell chamber. The cellular proliferation and cell cycle phase were examined respectively by MTT assay and flow cytometry analysis （FCM）. Results The siRNA against HPV16 E6 downregulated the expression of mRNA and protein dramatically in HNE-1 cells, with a rate of 66.3% and 56.7% respectively （P 〈 0. 05）. The invasion assay demonstrated that the number of HNE-1 cells which crossed the membrane in siRNA transfection group （45.3 ± 4.0） was significantly lower than that in negative control group （ 147.7 ± 4.7 ） （P 〈 0. 05 ）. After transfection, the proliferation of HNE-1 cells were inhibited and there was a remarkable difference compared with the control group （P〈0.05）. FCM showed that the cell cycle was arrested in the G0/G1 phase. Conclusion siRNA against HPV16 E6 gene effectively inhibits the specific gene expression, and then restrain the invasion and metastasis ability of HNE-1 cells, inhibits cell proliferation, and induces cell cycle to redistribute.

关 键 词：小干扰RNA 人乳头瘤病毒16 E6 侵袭 增殖 细胞周期 

分 类 号：R73-36[医药卫生—肿瘤] R730.23[医药卫生—临床医学]