野生型p53基因转染对人HCC-9204细胞端粒酶活性影响  

Influence of wild-type p53 gene tranfection on telomerase activity in human HCC-9204 cell

作　　者：戴大英[1] 翟为溶[1] 万大方[2] 朱腾方[1] 朱虹光[1] 

机构地区：[1]复旦大学上海医学院病理系,上海200032 [2]上海市肿瘤研究所癌基因及相关基因国家重点实验室,上海200032

出　　处：《中国癌症杂志》2003年第6期515-517,522,共4页China Oncology

摘　　要：目的 :探讨肿瘤抑制基因 p5 3对人肝癌细胞端粒酶活性及其催化亚单位hTERT (humantelomerasereversetranscriptase)的影响 ,并探讨可能的机制。方法 :用脂质体介导基因转染法将野生型p5 3(wt p5 3)导入端粒酶 /hTERTmRNA阳性、内源性 p5 3突变的HCC 92 0 4人肝癌细胞中 ;用Westernblot鉴定转染效率 ;PCR ELISA、TRAP 银染、原位杂交和RT PCR法检测细胞端粒酶活性和hTERTmRNA表达 ;用流式细胞仪观察细胞凋亡改变。结果 :外源wt p5 3基因转染后HCC 92 0 4细胞端粒酶活性明显受抑 ,hTERTmRNA表达水平下调 ,bcl 2表达减少 ,细胞呈现凋亡改变。结论 :外源wt p5 3基因表达可抑制人肝癌细胞端粒酶活性 ;端粒酶活化、hTERTmRNA表达存在 p5 3依赖性调节途径。Purpose:To investigate the effects of tumor suppressor gene p53 on the activity of telomerase/hTERT in human HCC-9204 cell line.Methods:Lipofection-mediated gene transfection method was used to transfect wild-type p53(wt-p53) gene into HCC 9204 cell which expresses telomerse /hTERT and carries mutant-type p53 gene. The expression of p53 was confirmed by Western blot. Both the telomerase activity and hTERT mRNA expression were detected by PCR-ELISA,TRAP-silver staining,in-situ hybridization and RT-PCR methods. The apoptotic appearance was examined by FCM.Results:Higher telomerase activity and hTERT mRNA level were detected in HCC-9204,and they were markedly inhibited after transfection with wt-p53. Meanwhile,decreasing level of bcl-2 protein and appearance of apoptosis were also shown in the transfected cells.Conclusions:Over expression of the exogenous wt-p53 gene does suppress both telomrease activity and hTERT mRNA expression in HCC cell line. There is a p53-dependent regulatory pathway for activation and expression of telomerase/hTERT in HCC.

关 键 词：肝细胞肝癌 肿瘤抑制基因P53 基因转染 端粒酶 

分 类 号：R73-362[医药卫生—肿瘤] R735.7[医药卫生—临床医学]