2006•Di-Si Junyi Daxue xuebaoRequires access
Effect of bcl-2 gene transfection on the proliferation of human gastric epithelial cell GES-1
Li Zhu
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Abstract
AIM:To observe the effect of bcl-2 gene transfection on the proliferation of human gastric epithelial cell GES-1. METHODS:The eukaryotic vector carrying the full length of bcl-2 gene and the neomycin resistance gene (pcDNA3/bcl-2) and the one only carrying the neomycin resistance gene (pcDNA3) were amplified and purified. The plasmid of pcDNA3/bcl-2 was used to transfect GES-1 cells by using the lipofectamine, and the empty vector pcDNA3 to transfect GES-1 cells as controls. And then, the cells were cultured in 1640 culture medium containing an appropriate concentration of G418 for selection. The method of HRP label immunohistochemical staining was used to identify the cells expressing bcl-2 gene positively. The morphologic changes of GES-1 cells were observed under an optical microscope by HE staining. The cell proliferation affected by the transfection of bcl-2 was measured by cell counting and MTT assay. RESULTS: bcl-2 gene and the vector pcDNA3 were transfected separately into GES-1 cells by lipofectamine. After G418 selection, the cells were transfected steadily. Compared with controls, HRP label immunohistochemical staining showed that bcl-2 gene transfected cells expressed Bcl-2 protein much more obviously. No morphologic changes were observed by HE staining. The growth curve was drawn by the method of cell counting,which showed that the growth of bcl-2 gene transfected cells was faster than that of empty vector transfected cells and untransfected cells after 6 d culture. And MTT assay showed that the growth of bcl-2 gene transfected cells were 4.15±0.31,5.98±0.56 and 8.94±0.79, respectively. While MTT assay showed that the growth of control cells were 3.01± 0.20 , 4.76±0.52 and 7.69±0.84, respectively. Significant differences were found between the 2 groups (P 0.05). CONCLUSION: The transfected of bcl-2 gene makes the cells express Bcl-2 protein steadily and highly, and enhances the cells' proliferation and malignancy.