The inhibitory effects on hepatitis B virus gene expression in stable expression of DN mutants of hepatitis B virus X gene
- VernacularTitle:HBx-GFP DN突变子长期稳定表达抑制2.2.15细胞乙型肝炎病毒基因的表达
- Author:
Jiawu SONG
;
Jusheng LIN
;
Xinjuan KONG
- Publication Type:Journal Article
- Keywords:
HBV X gene;
Dominant negative mutants;
Hepatitis B virus;
Gene therapy
- From:
Chinese Journal of Infectious Diseases
2001;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective The newly developing gene therapy method and dominant negative mutants were bein g used as new promising HBV therapy method, and a dominant negative mutant of HB V X g ene we have reported in our previous report has some effects both on HBV replica tion and expression in transient expression, but the effects were interfered by persistent secretion of HBV in HepG 2 2.2.15 cell line in the experiment. To mak e sure the effects of dominant negative mutant of HBx gene, we established a HBx DN stably expressing cell clone, and evaluated the effects of HBx dominant negat ive mutant on HBV gene expression. Methods The prev HBx-GFP dominant mutant and the plasmids pRev Xwt, pRev GFP which contain the wild type X gene or GFP gene then transfected into HepG 2 2.2.15 cells by liposome. The HBsAg, HBeAg by in media were as sayed by RIA and HBV-related RNA were assayed by Northern blot. Results The pRev HBx-GFP, GFP and wild type X constructs can be effectively expressed in HepG 2 2.2.15 cells. The stable expressed HBx -GFP can significantly reduce HBeAg, HBeAg in media and the HBV-related RNA in HepG 2 2.2.15 cells, but not for pRev Xwt and pRev GFP. Conclusions The dominant negative mutant pRev HBx-GFP can significantly inhibit the HBV gen e expression. It also suggested that X gene may be one promising target for HBV gene therapy.
