Effect on growth of human hepatoma cells by antisense gene of type Ⅱ insulin-like growth factor receptor transfection
- VernacularTitle:人胰岛素样因子Ⅱ型受体反义基因转染对肝癌细胞生长的影响
- Author:
Wensheng CHEN
;
Weiwen LIU
;
Jianren GU
- Publication Type:Journal Article
- Keywords:
IGF ⅡR;
Antisense gene;
Hepatoma cell
- From:
Chinese Journal of Digestion
1996;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the growth inhibition of human hepatoma cells tranfected with type Ⅱ insulin like growth factor receptor (IGF ⅡR) antisense gene. Methods We constructed the recombinant eucaryotic expression plasmids of IGF ⅡR sense and antisense genes, which were transfected into SMMC 7721 human hepatoma cell line using calcium phosphate coprecipitation, and selected in DMEM supplemented with 500 ?g/ml G418(Geneticin) for 2～3 weeks. Then the total number of colonies formed was counted. The effect of IGF ⅡR antisense gene transfection on endogenous IGF ⅡR mRNA levels was examined by Northern blot, and the growth rate and the ability of the transfected cells to form colonies in soft agar medium were also examined. Results The hepatoma cells transfected with IGF ⅡR antisense gene exhibited significant reduction in endogenous IGF ⅡR messenger RNA(mRNA) levels and loss of their ability for anchorage independent growth in soft agar. The IGF ⅡR antisense gene supressed the formation of colonies of SMMC 7721 cells resistant to G418 without alteration on cell growth curve. Conclusion The IGF ⅡR antisense gene expression effectively blocks IGF Ⅱ to IGF ⅡR autocrine and/or paracrine signal transduction pathway and reverses certain aspects of the cell malignant phenotype.
