MDA7基因的腺病毒载体构建及对肿瘤细胞凋亡的影响（临床医学论文）.docVIP

MDA7基因的腺病毒载体构建及对肿瘤细胞凋亡的影响（临床医学论文） 文档信息 属性： F-00BP33，doc格式，正文7241字。质优实惠，欢迎下载！ 适用： 作为医学论文、临床医学写作的参考文献，解决如何写好实用应用文、正确编写文案格式、内容摘取等相关工作。 作者： 王艳 目录 TOC \o 1-9 \h \z \u 目录 1 正文 1 搞要 1 关键字：复制缺陷型腺病毒 端粒酶启动子 肿瘤基因治疗 2 1 材料和方法 4 2 结 果 5 3 讨 论 7 【参考文献】 8 论文原创声明（模板） 9 论文致谢（模板） 10 正文 MDA7基因的腺病毒载体构建及对肿瘤细胞凋亡的影响（临床医学论文） 搞要 摘要：构建一种端粒酶启动子调控目的基因MDA?7/hIL24表达的复制缺陷型腺病毒，并初步探索其抗肿瘤活性。方法：通过基因操作技术将启动子（hTERTp）+目的基因(MDA?7)插入到5型腺病毒E1A区域，构建复制缺陷型腺病毒AdTP?mda7；同时构建对照病毒AdTP?EGFP。TCID50法测定病毒滴度。应用蛋白质印迹法检测MDA?7在肿瘤细胞株中的表达状态；通过结晶紫染色实验检测两种病毒对肿瘤细胞的杀伤差异。结果：成功构建携带目的基因的腺病毒载体，PCR鉴定正确；蛋白质印迹结果表明MDA?7选择性地在多种肿瘤细胞株中表达,诱导肿瘤细胞凋亡，如A549，SGC?7901等，在以相同MOI值感染原代成纤维细胞时，原代细胞中没有检测到MDA?7表达；结晶紫染色试验结果表明AdTP?mda7对肿瘤细胞株的杀伤能力明显高于对照病毒AdTP?EGFP。结论：成功构建复制缺陷型腺病毒AdTP?mda7，并证实MDA?7蛋白在肿瘤细胞中过表达诱导多种肿瘤细胞株发生凋亡。中国论文发表 关键字：复制缺陷型腺病毒 端粒酶启动子 肿瘤基因治疗 ［Abstract］ Objective: To develop a proliferation?deficient adenovirus for gene therapy in which the hTERT promoter was introduced to regulate the expression of the target gene in tumor ： A tumor?specific proliferation?difficient adenovirus vector was constructed by employing the human telomerase reverse transcriptase (hTERT)promoter to drive the expression of adenovirus?mediated target gene; at the same time AdSuTP?EGFP was constructed as was obtained by TCID50 expression of MDA?7 was detected by western blot and the appearance of senescence was assessed by violet were photographed 2h after staining. Results： Two adenovirus vectors including target gene were successfully constructed and identified by PCR blot analysed that MDA?7 selectively expressed in tumor cell lines MHCC, SGC7901, A549, and so on, and induced apoptosis，while primary lung fibroblast was transduced at equal MOI level and did not express detectable levels of MDA?7; violet staining test verified directly that killing effect of AdTP?mda7 was great stronger than AdTP?EGFP，supporting that one of the functions of MDA?7 was apoptosis induction. Conclusion： proliferation?deficient adenovirus, AdTP?mda7, was s