Construction a lentiviral vector for RNA interference of glutathione peroxidase 2 gene and its effect on HepG2 cell apoptosis.
- Author:
Jiangping CAO
1
;
Liujun TANG
;
Jianhong ZHANG
;
Yiqun ZHAN
;
Xiaoming YANG
;
Changhui GE
Author Information
- Publication Type:Journal Article
- MeSH: Apoptosis; Down-Regulation; Genetic Vectors; Glutathione Peroxidase; genetics; Hep G2 Cells; Humans; Lentivirus; RNA Interference; RNA, Small Interfering
- From: Journal of Southern Medical University 2015;35(6):832-837
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct a RNA interference lentiviral vector for human glutathione peroxidase 2 (GPX2) gene and observe the effect of GPX2 knockdown on cell apoptosis.
METHODSThe sequence of the small interfering RNA (siRNA) for GPX2 interference was inserted into the pSicoR vector. HepG2 cells were infected by the packaged si-GPX2 lentivirus and the expression of GPX2 in the infected cells was detected by both RT-PCR and Western blotting. Changes of cell apoptosis following the infection were analyzed by flow cytometry.
RESULTSThe lentiviral particles pSicoR-GPX2 were successfully packaged. The expression of GPX2 in the infected cells was obviously down-regulated at both RNA and protein levels. GPX2 knockdown caused increased apoptosis rate, increased Bax expression and lowered Bcl-2 expression in HepG2 cells.
CONCLUSIONWe have successfully constructed the lentiviral vector for RNA interference of human GPX2 gene.