Construction of eukaryotic expression plasmid of Smad-7 and study on its function as TGF-β1 inhibitor
- VernacularTitle:人肾系膜细胞Smad-7稳定表达细胞系的筛选及其拮抗TGF-β1作用
- Author:
Ping ZHOU
;
Jie LI
;
Jianzhen FANG
- Publication Type:Journal Article
- Keywords:
DNA-binding proteins;
Kidney;
Transforming growth factor beta
- From:
Journal of Chinese Physician
2008;10(7):873-875
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct eukaryctie expression vectors stably expressing Smad-7 and study its function as TGF-β1 inhibi- tor. Methods Smad-7 genes were obtained by PCR from fetal liver cDNA library and inserted in to eukaryotic expression plasmid pcD- NA3. 1/myc-his-B (-). The constructed plasmids containing right sequence of target genes were transfected into human mesangial cells by lipofectamine. The ceils were selected by G418, the expression of Smad-7 was detected by Western-blotting. Cell proliferation was detected by MTT method, and cell cycle was detected by Flow Cytometer to study its function as TGF-β1 inhibitor. Results We successfully con- struct Smad-7 expression plesmid. Smad-7 can inhibit the effects of TGF-[M induced GI cell cycle block and apoptosis. Conclusion The eukaryotic expression vector of Smad-7 has been successfully constructed. Over-expression of exogenous Smad-7 can inhibit the effects of TGF-β1 induced GI cell cycle block and apoptosis.
