腺病毒过表达及 siRNA 干扰人肝癌 HepG2细胞对11β-羟基类固醇脱氢酶 I表达的影响

摘要

Objective To design a adenovirus vector based system and specific siRNA for exploring the effects of overexpression and siRNA interference of the human hepatocellular carcinoma cell line HepG 2 on the ex-pression of 11β-hydroxysteroid dehydrogenase 1.Methods A adenovirus overexpression vector and specific siRNA targeting the 11β-HSD1 gene was designed and transfected into HepG 2 cells.Expression of 11β-HSD1 mRNA and protein was analyzed by real time PCR and Western blotting .Results A adenovirus transfection resulted in upgrade of 11β-HSD1 protein, and siRNA transfection led to inhibition of 11β-HSD1 mRNA and protein.Conclusion Trans-fection of the hepatocellular carcinoma cell line HepG 2 using a adenovirus vector and siRNA interference can signifi-cantly influence expression of 11β-HSD1 mRNA and protein .%目的：观察腺病毒（Ad-11β-HSD1）过表达系统及特异性siRNA（siRNA-11β-HSD1）干扰人肝癌HepG2细胞对11β-羟基类固醇脱氢酶I（11β-HSD1）表达的影响。方法应用Ad-11β-HSD1以及siRNA-11β-HSD1转染人肝癌HepG2细胞，用实时荧光定量PCR及Western Blot 检测11β-HSD1表达。结果转染72 h后，对照组及腺病毒过表达组细胞中均可见明显 GFP荧光，且转染48 h后蛋白水平明显升高；siRNA干扰48 h后，mRNA及蛋白水平均明显降低。结论腺病毒转染可有效过表达11β-HSD1，而siRNA干扰可有效抑制11β-HSD1表达。