Expression Of Survivin And The Affection Of Its Antisense Gene On Cell Proliferation, Differentiation And Apoptosis In Glioma

Objective: To investigate the protein expression of survivin and the affection of its antisense gene on cell proliferation ,differentiation and apoptosis in glioma so as to provide basic data for further study on the roles of surviving in the development of glioma.Methods:1. The immunohistochemistry was used to measure protein expression of survivin and PCNA, and TUNEL was used to detect cell apoptosis in 91 cases of glioma with gradeâ… 26 cases, grade â…¡ 54 cases, grade â…¢-â…£ 11 cases. 2. Molecular cloning technique was used to construct the antisense survivin eukaryotic expression vector pIRES2-EGFP-SVVas .And then the pIRES2-EGFP-SVVas was transfected into SHG-44 cells with lipofectamine. The expression of surviving gene on the transfected cells was measured by RT-PCR and western blot . The protein expression of GFAP and PCNA was measured by immunohistochemistry, the cell growth curve by MTT , cell cycle by FCM, and cell apoptosis by TUNEL.Results:1. The expression of survivin was positive in glioma and it was positively relative with malignancy of glioma. The positive percent of survivin was 30.8% with glioma gradeâ… , 57.4% with gradeâ…¡,and 72.7% with grade â…¢-â…£, respectively. 2.The expression of survivin was negatively relative with apoptosis index and positively relative with labeling index of PCNA in glioma. 3. The pIRES2-EGFP-SVVas vector was constructed successfully and transfected into SHG-44 cells by lipofectamine.The expression of survivin gene was found decreasing in the cells transfected with the pIRES2-EGFP-SVVas. 4. The cells transfected with pIRES2-EGFP -SVVas grew slower than those transfected with pIRES2-EGFP only and SHG-44 cell with no transfected. Cell cycle examination showed that the proportion of cells increased in G0/G1 phase and decreased in G2/M phase and S phase period in the cells transfected with pIRES2-EGFP-SVVas compared with those transfected with pIRES2-EGFP. The cells transfected with pIRES2-EGFP-SVVas was higher AI and lower LI than those transfected with pIRES2-EGFP only.Conclusions:1. The expression of survivin was positive in glioma and it was positively relative with malignancy of glioma. 2. The pIRES2-EGFP-SVVas vector was constructed successfully and transfected into SHG-44 cells by lipofectamine. 3. The pIRES2-EGFP-SVVas can decrease the expression of survivin gene on the cell transfected with the vector . 4. Stable transfection with antisense survivin inhibited cell growth , proliferation,and promoted cell differentiation and apoptosis.