Effects of rhTGF-?_1 and TGF-?_1 gene transfection on the proliferation of cultured rabbit corneal endothelial cells in vitro
- VernacularTitle:rhTGF-?_1及转染TGF-?_1基因对兔角膜内皮细胞增殖的影响
- Author:
Huixiang MA
;
Jintang XU
;
Zhenyou JIANG
;
Suimei ZHANG
;
Songbin ZHAO
;
Jiansu CHEN
- Publication Type:Journal Article
- Keywords:
Transforming growth factor beta;
Gene transfection;
Corneal endothelial cells;
Cell proliferation
- From:
Chinese Journal of Pathophysiology
1989;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To investigate the effects of rhTGF-?1 and TGF-?1 gene transfection on the proliferation of cultured rabbit corneal endothelial cells in vitro.METHODS:Cell growth induced by various concentrations of rhTGF-?1 was determined by MTT proliferation assay.Under the induction of liposomes,recombinant pSecTag2-TGF-?1MP vectors were transferred into the corneal endothelial cells.Morphological changes of transfected cells were observed by HE staining.The expression levels of TGF-?1 were assessed by ELISA.Cell cycle analysis was assessed by flow cytometry.DNA fragment analysis was used to confirm the presence of apoptosis.RESULTS:rhTGF-?1 in concentrations of 5-20 ?g/L showed a significant suppressive effect on the proliferation of corneal endothelial cells,0.5-1 ?g/L had no effect,0.05-0.1 ?g/L facilitated cell growth,as compared with negative controls.The morphous of transfected corneal cells had no significant abnormality compared with normal cells.According to the result of ELISA,the concentration of TGF-?1 in the supernatant was calculated to be(98?3)ng/L.Flow cytometry assay showed that S and G2/M phase of transfected cells decreased significantly compared with that of control group,but the cell cycle recovered normally after adding 10 ?g/L EGF into the culture medium.Agarose electrophoresis didn't show marked ladders in transfected group.CONCLUSION:Effects of rhTGF-?1 on the proliferation of corneal endothelial cells are different with various concentrations.TGF-?1 gene transfection shows suppressive effect on the proliferation of cultured corneal endothelial cells,but does not induce cell apoptosis.EGF is the antagonist of this suppressive effect.
