中文摘要：

目的：构建hTERT启动子调控的肿瘤坏死因子相关的凋亡诱导配体（TRAIL）基因表达载体,探讨其对肝癌细胞SMMC7721和肝细胞HL7702增殖的抑制作用。方法：采用分子生物学方法构建重组质粒pshuttle-TRAIL和pshuttle-hTERT-TRAIL,经PCR和酶切鉴定正确后,通过脂质体转染SMMC7721和HL7702细胞,同时转染pcDNA3.1＋-GFP并用荧光显微镜检测转染效率。MTT法检测重组质粒对细胞增殖变化的影响。结果：成功构建重组质粒pshuttle-TRAIL和pshuttle-hTERT-TRAIL,荧光显微镜检测结果显示：质粒与脂质体的比例为1∶3时细胞转染效率最高。pshuttle-hTERT-TRAIL组SMMC7721细胞增殖抑制率从16h开始明显增加,与对照组比较差异有显著性（P〈0.001）,且从24h开始与pshuttle-TRAIL组比较细胞增殖抑制率明显增加（P〈0.05）;但重组质粒对HL7702细胞增殖抑制率无影响。结论：hTERT启动子调控的TRAIL可明显抑制肝癌细胞的增殖,而对人肝细胞的增殖无影响,其作用效果明显优于TRAIL单基因治疗。

英文摘要：

Objective To construct the hTERT promoter-driven TRAIL expression vector and explore its inhibitory effects on the proliferation of hepatoma SMMC7721cells and HL7702hepatocytes.Methods The recombinant plasmids pshuttle-TRAIL and pshuttle-hTERT-TRAIL were constructed with molecular biological methods.They were confirmed correctly by endonucleases digestion and PCR identification,and transfected into SMMC7721and HL7702cells through liposome,meanwhile,the pcDNA3.1＋-GFP was transfected to detect the transfection efficacy by fluorescence microscopy.MTT was used to detect the effects of the recombinant plasmids on cell proliferation in transfected cells.Results The recombinant plasmid pshuttle-TRAIL and pshuttle-hTERT-TRAIL were constructed successfully.And the transfection efficacy was highest by fluorescence microscopy when the ratio of plasmid to liposome was 1∶3.As compared with control group,the inhibitory rates of SMMC7721cells from 16hafter transfection in pshuttle-hTERT-TRAIL group was increased significantly（P〈0.001）;and as compared with pshuttle-TRAIL group,the inhibitory rates of SMMC7721cells from 24hafter transfection in pshuttle-hTERT-TRAIL group was increased significantly（P〈0.05）.However,as compared with control group,the inhibitory rates of HL7702cells in recombinant plasmids groups did not change significantly.Conclusion hTERT promoter-driven TRAIL could significantly inhibit the hepatoma cell proliferation,especially superior to that in pshuttle-TRAIL,however,no effects on hepatocyte proliferation.