短发夹RNA介导的RNA干扰技术在HepG2.2.15细胞中的应用研究  

The role of shRNA-mediated RNA interference in HepG2.2.15 cells:an application study

作　　者：杨慧[1] 赵龙凤[1] 赵中夫[2] 张国英[2] 刘明社[2] 

机构地区：[1]山西医科大学第一医院感染病科,太原030001 [2]长治医学院肝病研究所

出　　处：《中国药物与临床》2011年第5期502-504,I0001,共4页Chinese Remedies & Clinics

基　　金：山西省自然科学基金(20051114)

摘　　要：目的探讨shRNA真核表达质粒介导的RNA干扰技术对HepG2.2.15肝癌细胞中乙型肝炎病毒(HBV)复制及甲胎蛋白(AFP)表达的特异性抑制作用。方法采用时间分辨荧光免疫测定技术(TRFIA)检测乙型肝炎表面抗原(HBsAg)和乙型肝炎e抗原(HBeAg)含量;采用微粒子化学发光免疫分析法(MLFA)检测甲胎蛋白含量。采用免疫细胞化学技术检测细胞中HBsAg及甲胎蛋白的表达。结果 pGenesil-shHBV X对HepG2.2.15细胞HBsAg、HBeAg的特异性抑制作用随时间而递增(P<0.05),pGenesil-shAFP对HepG2.2.15细胞甲胎蛋白的特异性抑制作用同样随时间而递增(P<0.05),且二者之间无交叉抑制作用。结论 shRNA真核表达质粒pGene-sil-shHBV X介导的RNA干扰作用可特异性抑制HepG2.2.15细胞HBV的复制表达,pGenesil-shAFP可特异性抑制HepG2.2.15细胞甲胎蛋白的表达,为RNA干扰技术抗病毒及抑制肿瘤的进一步研究奠定基础。Objective To explore the specific inhibitory effect of shRNA eukaryotic expression plasmids mediated RNA interference on HBV virus replication and AFP protein expression in HepG2.2.15 hepatoma carcinoma cells（HCCs） . Methods The contents of HBsAg and HBeAg were detected by time-resolved immunofluorometric assay kit（TRFIA） ,the content of AFP by microparticle chemicluminescence immunoassay（MCIA） . And the expression of HBsAg and AFP proteins by immunohistochemistry. Results The specific inhibitory effect of pGenesil-shHBV X on HBsAg and HBeAg in HepG2.2.15 cells increased along with the advance of time（P0.05） ,and that of pGenesil-shAFP on AFP in HepG2.2.15 cells also increased along with the increase of time（P0.05） . Moreover,no cross-inhibitory effect was found between pGenesil-shHBV X and pGenesil-shAFP. Conclusion The shRNA eukaryotic expression plasmids pGenesil-shHBV X mediated RNA interference could specifically inhibit HBV replication,while pGenesil-shAFP could inhibit AFP expression in HepG2.2.15 cells,which laid the foundation for further studies of RNA interference technology to resist virus and inhibit tumor.

关 键 词：RNA干扰 肝炎病毒 乙型 甲胎蛋白 SHRNA HEPG2.2.15 

分 类 号：R735.7[医药卫生—肿瘤]