特异性siRNA抑制HPV16 E7基因表达的实验研究  

Specific siRNA inhibits HPV16 E7 gene expression: an experimental study

作　　者：张晓[1] 王传新[1] 李伟[1] 郑桂喜[1] 张建[1] 阚士锋[1] 王立水[1] 

机构地区：[1]山东大学齐鲁医院检验科,济南250012

出　　处：《山东大学学报（医学版）》2009年第11期64-67,共4页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金资助项目(30672010)

摘　　要：目的筛选针对HPV16 E7基因有效的siRNA,探讨其对HaCaT-E7细胞中HPV16 E7 mRNA及细胞表面HLA-Ⅰ类分子表达的影响。方法采用化学法合成3条HPV16 E7特异性siRNA,应用转染试剂Lipofectamine2000将其转染入HaCaT-E7细胞,采用实时荧光定量PCR(RT-PCR)检测HPV16 E7 mRNA的表达,采用流式细胞术(FCM)检测细胞表面HLA-Ⅰ类分子的表达。结果3条siRNA均能有效抑制HaCaT-E7细胞中HPV16 E7的转录表达,以siR-NA2作用效果最明显,抑制率达75%,细胞膜表面HLA-Ⅰ类分子的表达明显上调,平均荧光强度(MFI)为130.18±1.07,高于空转染对照组(100.32±3.01)和非特异性siRNA对照组(100.82±2.87)。结论化学合成的HPV16 E7特异性siRNA能有效抑制HaCaT-E7细胞中E7 mRNA的表达,同时使细胞表面HLA-Ⅰ类分子表达上调。Objective To screen an effective HPV16 E7 specific siRNA and investigate its effect on expression of HPV16 E7 mRNA and cell surface HLA class I antigen in HaCaT-E7 cells.Methods Three HPV16 E7 specific siRNAs were chemically synthesized and transfected into HaCaT-E7 cells by Lipofectamine2000.Expression of HPV16 E7 mRNA was determined by real-time PCR while expression of cell surface HLA class I antigen was determined by flow cytometry（FCM）.Results All three siRNAs effectively inhibited expression of HPV16 E7 mRNA in HaCaT-E7 cells.Of the siRNAs,siRNA2 was the most effective and its inhibitory rate reached 75%.Expression of cell surface HLA class I antigen was obviously up-regulated.Post-transfected average mean fluorescence intensity（MFI） of HaCaT-E7 cell surface HLA class I antigen was 130.18±1.07,which was significantly higher than that in the mock control group（100.32±3.01） and the non-specific siRNA control group（100.82±2.87）.Conclusion HPV16 E7 specific siRNA can effectively inhibit expression of E7 mRNA in HaCaT-E7 cells and induces up-regulation of cell surface HLA class I antigen.

关 键 词：乳头状瘤病毒 人 HLA抗原 基因 RNA干扰 HACAT细胞 

分 类 号：R737.33[医药卫生—肿瘤]