Qu Jian-Hui, Zhu Ming-Hua, Lin Jing, Ni Can-Rong, Li Fang-Mei, Zhu Zhi, Yu Guan-Zhen
Department of Pathology, the Second Military Medical University, Shanghai 200433, China.
World J Gastroenterol. 2005 Oct 21;11(39):6212-5. doi: 10.3748/wjg.v11.i39.6212.
To investigate the contribution of HBV in the development of hepatocarcinoma by examining the effects of HBV on p53 function in SMMU-7721 cell line.
Plasmid pCMVp53 was transfected or cotransfected with pCMVHBVa (wild-type HBV) or PCMVHBVb (mutation type HBV) into the hepatoma cell line SMMU-7721 by lipofectamine. Apoptosis cells were labeled with annexin V-FITC and confirmed by flow cytometry. Reporter plasmid PG13-CAT or p21-luc was cotransfected, respectively, into each group to determine the transactivation activity of p53 and its effect on p21 promoter. Western blot was performed to observe p53 expression in hepatoma cell line of each group.
The group transfected with pCMVp53 alone exhibited higher luciferase activity and higher apoptosis rate, otherwise, the p53 expression and reporter activity of PG13-CAT or P21-luc as well as cell apoptosis rate were obviously higher in the group cotransfected of pCMVp53 with pCMVHBVa, but not in the other cotransfected group.
Transient transfection of HBV into the SMMU-7721 cell line can enhance p53 expression and its effects on development of hepatocarcinoma.
通过检测乙肝病毒（HBV）对SMMU - 7721细胞系中p53功能的影响，探讨HBV在肝癌发生发展中的作用。
采用脂质体法将质粒pCMVp53分别与pCMVHBVa（野生型HBV）或PCMVHBVb（突变型HBV）共转染至肝癌细胞系SMMU - 7721。用膜联蛋白V - FITC标记凋亡细胞，通过流式细胞术进行鉴定。将报告质粒PG13 - CAT或p21 - luc分别共转染至各组，以检测p53的反式激活活性及其对p21启动子的影响。采用蛋白质免疫印迹法观察各组肝癌细胞系中p53的表达情况。
单独转染pCMVp53的组显示出较高的荧光素酶活性和凋亡率。此外，pCMVp53与pCMVHBVa共转染组的p53表达、PG13 - CAT或P21 - luc的报告活性以及细胞凋亡率均明显高于其他共转染组。
将HBV瞬时转染至SMMU - 7721细胞系可增强p53表达及其对肝癌发生发展的影响。
