The construction of siRNA-tTG lentivirus vector and its effect on activated hepatic stellate cells.
- Author:
Gang ZHAO
1
;
Zhi-Qi ZHANG
;
Zhi-Yong WU
;
Bo CHEN
;
Bin ZHANG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cell Line; Genetic Vectors; Hepatic Stellate Cells; Lentivirus; RNA, Small Interfering; genetics; Rats; Transfection; Transglutaminases; genetics
- From: Chinese Journal of Surgery 2009;47(9):701-704
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct the lentivirus vector of siRNA-tTG and investigate the effects and mechanism of lentivirus mediated tTG RNA interference (RNAi) on the hepatic stellate cell (HSC).
METHODSThe effective RNAi sequences targeted the tTG gene were designed and Lenti-siRNA-tTG was constructed by plasmid transfection. The rats were distributed into three groups:negative control (CON) group, siRNA-GFP (GFP) group and siRNA-tTG (rtTG) group. After the infection of lentivirus mediated siRNA-tTG, the mRNA and protein expression of tTG, alpha-SMA and collagen-1, and the cell proliferative status of HSC were observed in the three groups respectively.
RESULTSThe lentivirus vector of siRNA-tTG was constructed successfully. After the infection of lentivirus mediated siRNA-tTG, the mRNA expression of tTG and collagen-1 was decreased significantly (P < 0.05), and the protein expression of tTG, lysine and collagen-1 was also decreased significantly (P < 0.01), in the same time the proliferative ability of HSC was decreased significantly (P < 0.05).
CONCLUSIONSLentivirus mediated siRNA-tTG can inhibit the proliferation of HSC by inhibition of the expression of tTG and attenuate the synthesis of collagen-1 and lysine. Thus, it may have a potential anti-fibrosis effect.