Effect of siRNA on hepatitis B virus X gene to HepG2.2.15 cells
10.3969/j.issn.1006-5725.2016.13.013
- VernacularTitle:乙型肝炎病毒X基因siRNA对HepG2．2．15细胞的影响
- Author:
Jinhong ZHU
;
Chunmiao WANG
;
Hong WU
;
Meishuang LIN
;
Xuelian ZHU
;
Maofeng WU
;
Xiaoman WU
- Publication Type:Journal Article
- Keywords:
RNA interference;
HepG2.2.15;
Hepatitis B virus;
Transfection
- From:
The Journal of Practical Medicine
2016;32(13):2120-2123
- CountryChina
- Language:Chinese
-
Abstract:
Objective Using chemically synthesized small interfering RNA (siRNA) transfected HepG2.2.15 cells to construct a cell model in interfering hepatitis B virus (HBV) X gene, studying the inhibi-tion of HBV replication and antigen expression in vitro. Methods After transfection of HepG2.2.15 cell for 24 h, 48 h, 72 h, detecting the cell supernatant of HBsAg and HBeAg by chemiluminescence immunoassay, the cell supernatant HBxAg protein by ELISA , the HBx mRNA relative expression of transfected cell was detected by fluorescence quantitative polymerase chain reaction (PCR), the ability of cell proliferation was detected by CCK-8 assay. Results After HBx-siRNA transfected HepG2.2.15 cells, cell proliferation ability was inhibited. The cell of HBx mRNA and the cell supernatant of HBxAg expression decreased (P < 0.05); at the same time it in-hibited the expression of HBsAg and HBeAg. The suppressed peak and the inhibition rate were 66% and 58%respectively at 72 h. The fluorescence quantitative PCR confirmed that expression of HBV DNA in the super-natant was decreased. Conclusion The HepG2.2.15 cell interference model of HBV X gene has been success-fully constructed, which has an effect of inhibiting proliferation of HepG2.2.15 cells and replication and expres-sion of HBV gene in vitro.
