摘要
目的探讨ＨＢｘ蛋白对肝细胞Ｆａｓ配基（ＦａｓＬ）表达的调控作用。方法构建ＨＢｘ基因的真核表达载体ｐＣＥＰ４ｘ，电击法转染ＨｅｐＧ２细胞，潮霉素筛选阳性克隆细胞ＨｅＰＧ２Ｘ。用免疫细胞化学染色和免疫蛋白印迹法检测转染ＨＢｘ基因的ＨｅｎＧ２ｘ细胞ＦａｓＬ蛋白的表达。结果逆转录ＰＣＲ祛分析表明，ＨＢｘ基因在转染细胞内可有效转录；ＨｅｐＧ２ｘ细胞出现新的ＦａｓＬ蛋白表达。结论乙肝病毒ｘ蛋白可激活肝细胞表达ＦａｓＬ。
Objective To Study the effed of hepatitis B virus x (HBx) protein on the expression ofFas Ligsnd in HepG2 cells. Methods The open reading frame of HBx gene was obtained from HBVgrnome (subtype adw) digested with Nco Ⅰ and Bgl Ⅱ, then cloned into PCEP4. The reconstructive PlasmidpCEP4x was transfected into HepG2 cells by electroportion method. After selecting with hygfomycin B,resistant cloutes were obtained. The expressions of HBx gene and Fas Ligand gene were detected by RTPCR, immunohistochemical staining and western blot analysis. Results It showed that Fas Lignnd geneexpressed in the modified HepG2 cells transfected with plasmid pCEP4x. Conclusions The results suggestthe expression of Fas Lignnd in HepG2 cells could be activated by HBx protein.
出处
《中华肝脏病杂志》
CAS
CSCD
1999年第2期67-70,共4页
Chinese Journal of Hepatology
基金
本课题受国家自然科学基金!367634
卫生部科学基金
