人MUC4启动子驱动的HSV-TK重组腺病毒靶向杀伤SGC-7901胃癌细胞的实验研究  

Targeting cytotoxic effect of recombinant adenovirus carrying HSV-TK driven by MUC4 promoter on SGC-7901 gastric cancer cells

作　　者：陈衍[1] 韩晟 刘文超[1] 

机构地区：[1]第四军医大学西京医院肿瘤内科,陕西西安710032 [2]第四军医大学口腔医院信息科,陕西西安710032

出　　处：《中华肿瘤防治杂志》2012年第1期10-14,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(81001180);陕西省自然科学基金(2010JM4005)

摘　　要：目的:构建人粘蛋白(MUC4)启动子驱动下的单纯疱疹胸苷激酶基因(HSV-TK)重组腺病毒,研究其对SGC-7901胃癌细胞的靶向杀伤作用。方法:克隆MUC4启动子区625bp活性序列,构建重组荧光素酶报告基因载体pGL3-MUC4,检测其在SGC-7901胃癌细胞及NIH3T3成纤维细胞中的转录活性。以AdEasyTM腺病毒系统为载体,构建MUC4启动子驱动下的HSV-TK重组腺病毒rAdeno-MUC4-TK,感染SGC-7901及NIH3T3,经更昔洛韦(GCV)处理,MTT法检测细胞活力,TUNEL法检测细胞凋亡。结果:成功扩增出大小为625bp的MUC4启动子序列。pGL3-MUC4在SGC-7901细胞中具有强转录活性,转录活性高于强启动子SV40 6.6倍,而NIH3T3成纤维细胞系中几乎无转录活性。构建重组腺病毒rAdeno-MUC4-TK,MTT法和TUNEL检测结果显示,其与GCV联合能够诱导SGC-7901细胞凋亡,产生靶向细胞毒作用。结论:人MUC4启动子驱动下的HSV-TK重组腺病毒联合GCV对SGC-7901胃癌细胞具有靶向杀伤作用,MUC4启动子可以作为胃癌靶向基因治疗的工具。OBJECTIVE： To construct the recombinant adenovirus carrying herpes simplex virus thymidine kinase （HSV-TK） driven by human mucin 4 （MUC4） promoter and analyze its selective cytotoxic effect in vitro on human gas tric cancer cell line SGC 7901 combined with ganciclovir （GCV）. METHODS： The fragment of MUC4 promoter from hu man genome DNA and construct pGL3-MUC4 recombinant luciferase report vector were coloned and its transcriptional ac- tivities in SGC-7901 and NIH3T3 cell lines was analyzed. The recombinant adenovirus carrying HSV-TK driven by hu man MUC4 promoter （rAdeno-MUC4-TK） based on AdEasyTM adenoviral system was constructed and the selective cy- totoxie effect of the recombinant adenovirus on SGC-7901 cells combined with prodrug GCV was analyzed by MTT and TUNEL assays. RESULTS： Totally 625 bp MUC4 promoter fragment was coloned by PCR. pGI.3--MUC4 showed high tran- scriptional activitiy in SGC 7901 cell 6.6 time stronger than SV40 strong promoter but little in NIH3T3 fibroblast cell. rAdeno- MUC4-TK combined with C^V could evidently inhibit the proliferation of SGC-7901 cells and induce cell apoptosis, but showed low activity in NIH3T3 fibroblast cell. CONCLUSION： Adenoviral-mediated suicide gene therapy controlled by MUC4 promoter can induce specific cytotoxic effect on SCA2-7901 gastric cancer cells,suggesting MUC4 promoter can be serve as a useful tool for transcriptional targeting of gastric cancer gene therapy.

关 键 词：粘蛋白 启动区/遗传学 转录 遗传 基因疗法 腺病毒 人 

分 类 号：R735.2[医药卫生—肿瘤]