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Expression of Bmi1 in Hepatocellular Carcinoma and Hepatic Tumor Cell Line HepG2 and Their Relationship with Cell Proliferation and Apoptosis

Author: DangZheng
Tutor: ZuoKeFeng；SongWenJie
School: Fourth Military Medical University
Course: Surgery
Keywords: Hepatocellular carcinoma Bmi1 Cell proliferation Apoptosis Immunohistochemistry Flow cytometry
CLC: R735.7
Type: Master's thesis
Year: 2011
Downloads: 51
Quote: 0
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Abstract


Objective: To detect the expression of Bmi1 in HCC tissues and hepatoma cell lines , to explore the relationship between of Bmi1 HCC proliferation and apoptosis , and provides a new way of thinking for liver cancer diagnosis, assessment , treatment and prognosis . Method: apoptosis using immunohistochemical EnVision method to detect 54 cases of HCC specimens and corresponding adjacent tissues and 10 cases of normal liver tissue samples the Bmi1 gene and Ki67 antigen expression and TUNEL assay above specimens . Will be constructed pCMV6-Bmi1 vector transfected hepatoma cell line HepG2 were identified using Western blot technology and real-time quantitative PCR Bmi1 expression after 48 hours . And cell cycle and apoptosis of transfected cells using flow cytometry . Results: immunohistochemical and apoptosis detection HCC in of Bmi1 the high expression with age , gender , tumor size , number of tumors , clinical TNM stage cirrhosis and HBsAg infection was not significantly related ( P gt ; 0.05) , but with the histological grade , high in the differentiation group Bmi1 expression rate was significantly higher than that in poorly differentiated group ( P lt; 0.05 ) . Liver cancer Bmi1 positive group proliferation index ( PI ) ( 50.3 ± 21.4 ) % was significantly higher than the negative expression group ( 17.3 ± 7.1) % ( P lt; 0.05 ) , the apoptosis index (AI) , no significant differences ( P gt ; 0.05) . Western blot and quantitative real-time PCR results suggest that Bmi1 protein of Bmi1 mRNA overexpression in cell lines , and its growth curve visible the pCMV6-Bmi1 transfection of HepG2 cells proliferation rate than untransfected group and pCMV6 empty vector transfected group cells were significantly elevated . The flow cytometry results suggest that the pCMV6-Bmi1 transfection group pCMV6 empty vector transfected group , G2% S% significantly higher increase in DNA synthesis . The role of 5-FU 24h, apoptosis was detected , pCMV6-Bmi1 transfection group apoptosis rate was significantly lower than pCMV6 empty vector transfected cells . Conclusion: The above experiments suggest that Bmi1 involved in the regulation of liver cancer cell proliferation , promotes liver cancer early and cell proliferation, and the relationship between apoptosis could not be confirmed by experiment , the inference may be less relevant with the sample size of the study , or is not a direct regulation of the relationship , but multiple factors involved in the process , to await further experiments and research .

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Liver tumors
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