端粒酶逆转录酶基因启动子调控FADD表达载体诱导人结肠细胞凋亡的研究  

Expression of FADD Driven by hTERT promoter induces apoptosis of human colorectal carcinoma cells

作　　者：张险峰[1] 江应安[1] 印安宁[1] 罗和生[1] 王卫星[2] 

机构地区：[1]武汉大学人民医院消化内科,430060 [2]武汉大学人民医院普通外科,430060

出　　处：《中华实验外科杂志》2011年第7期1139-1141,F0004,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30471690）

摘　　要：目的构建人端粒酶逆转录酶（hTERT）基因启动子调控Fas相关死亡结构域蛋白（FADD）肿瘤细胞特异性表达载体，观察其对人结肠癌细胞凋亡的作用。方法利用hTERT基因核心启动子和FADD基因片段，构建hTERT基因启动子调控FADD的肿瘤细胞特异性表达载体。用脂质体转染法，将其分别转染人结肠癌细胞和正常细胞，观察人结肠癌细胞和正常细胞的端粒酶活性、细胞周期及凋亡。结果hTERT基因核心启动子调控FADD表达载体，在所检测的肿瘤细胞中具有明显的转录活性；而在正常细胞中则无明显的转录活性。Colon320细胞、LoVo细胞、SW480细胞与WI-38细胞凋广率在不同转染时间比较，差异有统计学意义（P均〈0．01）。WI-38转染pLNCX-hTERT—FADD与pLNCX—hTERT—GFP在24、48、72、96、120h凋亡率比较，差异均无统计学意义（P均〉0．05）。结论hTRET基因核心启动子具有肿瘤特异性，构建hTERT基因核心启动子调控FADD表达载体可能是一种新的肿瘤治疗途径。Objective To construct the tumor cell-specific expression vector of FADD expression driven by the haman telomerase reverse transeriptase （hTERT） gene promoter and observe its effects on apoptosis in vitro. Methods hTERT gene promoter and FADD gene were used to construct the tumor cell-specific expression vector of FADD expression driven by hTERT promoter. The vector was transfected the tumor cells and normal cells using LipofectamineTM 2000, and apoptosis of the human colonic carcinoma ceils was observed. Results FADD expression driven by the hTERT gene promoter exhibited apparent transcriptional activity in tumor cells but not in normal cells. There was significant difference in cell apoptosis rate among Colon320 cells, LoVo cells, SW480 cells and WI-38 cells at different transfection time points （P 〈 0. 01 ）. After transfection of pLNCX-hTERT-FADD and pLNCX-hTERT-GFP into WI-38 cells, there was no difference in apoptosis rate in 24, 48, 72, 96 and 120 h （ P 〉 0. 05 ）. Conclusion As hTERT gene promoter demonstrates tumor specificity, the expression veetor of FADD expression driven by hTERT gene promoter may provide a new approach to tumor therapy.

关 键 词：结肠癌 端粒酶逆转录酶基因启动子 基因治疗 FAS 

分 类 号：R735.35[医药卫生—肿瘤]