Construction of shSet7/9 vector and its function in HepG2
10.7652/jdyxb201506008
- VernacularTitle:sh-Set7/9表达载体的构建及其在 HepG2细胞中的功能
- Author:
Kejun MA
;
Xingchen SHI
;
Ping LI
;
Xiaoqiang LI
;
Wen REN
;
Long QIN
;
Xinhe SHI
- Publication Type:Journal Article
- Keywords:
Set7/9;
HepG2 cell;
Western blot;
methyltransferase;
vector construction;
hepatocellular carci-noma
- From:
Journal of Xi'an Jiaotong University(Medical Sciences)
2015;(6):753-757
- CountryChina
- Language:Chinese
-
Abstract:
Objective To silence human gene Set7/9 and screen out stable transfection cell line in hepatocellular carcinoma cell line HepG2 so as to investigate the impact of down-regulation of Set7/9 in cell line HepG2 and provide experimental foundation for studies on the effect of set7/9 in HepG2.Methods The target oligo was designed and synthesized;shRNA interference vector and the control vector were constructed and transfected into HepG2 cells;the stable transfection cells were screened out.Then Real-time PCR and Western blot were performed to detect the silence of Set7/9 according to both gene expression and protein expression level. Results The shRNA interference vector was constructed and transfected into HepG2 cells successfully.Compared with that in the negative control group,the expression of Set7/9 was dramatically downregulated (P < 0.05 ). Meanwhile,the expression of related protein Sirt1 and Suv39h1 was upregulated 8.4 folds and 1.1 fold, respectively.Conclusion Downregulation of Set7/9 expression can upregulate Sirt1 and Suv39h1,suggesting that Set7/9 may affect the activity of HepG2 cell lines.
