靶向Wnt/β-catenin通路的溶瘤腺病毒携带抑癌基因TSLC1抑制肿瘤细胞增殖的研究  

The Study on Inhibition of Tumor Cell Proliferation by Oncolytic Adenoviruses Targeting Wnt/β-catenin Pathway and Delivering Tumor Suppressor TSLC1

作　　者：金槿 赵雨佳[1] 章健[1] 王彬蓉 王毅刚[1] 

机构地区：[1]浙江理工大学生命科学学院,杭州310018

出　　处：《中国细胞生物学学报》2017年第11期1407-1414,共8页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81272687);浙江省自然科学基金(批准号:LY16H160056);浙江理工大学521骨干人才项目资助的课题~~

摘　　要：溶瘤病毒(oncolytic viruses)可选择性地感染肿瘤细胞并在其中复制,使宿主细胞裂解并在体内产生强烈的免疫应答反应,从而抑制或者杀死肿瘤细胞。因其特有的溶瘤性和靶向性,溶瘤病毒成为肿瘤基因治疗的热门领域之一。基于Wnt信号的肿瘤特异性,本研究使用Wnt信号启动子TCF/LEF结合位点序列,调控腺病毒早期基因E1A,并删除E1A基因序列上能与Rb蛋白结合的24 bp片段。将抑癌基因TSLC1插入腺病毒的E3区,形成重组腺病毒Ad.wnt-E1A(Δ24)-TSLC1。通过双荧光素酶报告系统、Western blot实验、MTT法、结晶紫染色法、Hoechst染色实验分别检测Wnt信号活性、肿瘤细胞的存活率和凋亡的变化情况。结果发现,肿瘤细胞较正常肝细胞具有较高的Wnt通路活性和病毒敏感性,其中重组病毒处理Hep G2后杀伤效果显著,并可通过Caspase通路诱导细胞凋亡,为肝癌的临床治疗提供参考。Oncolytic viruses are able to selectively infect and replicate in tumor cells, which further lyses tumor cells and produces immunological response to inhibit or kill tumor cells. Oncolytic viruses become one of the most promising fields for tumor gene therapy due to their special oncolysis and tumor targeting ability. Based on the tumor specificity of Writ signal pathway, our study used the novel constructed oncolytic adenovirus Ad.wnt- E1A（A24）-TSLC1 with a mutant E1A expression cassette driven by TCF/LEF binding site promoter and delivering a tumor suppressor gene TSLC1. The Writ signalling activity, survival rate and apoptosis of tumor cells are detected by Dual-Luciferase reporter, Western blot, MTT, crystal violet and Hoechst staining assay, respectively. The results show that tumor cells have higher Wnt signal activity and stronger sensitivity to oncolytic viruses comparing with normal cells. Ad.wnt-E1A（A24）-TSLC 1 can effectively kill HepG2 cells and induce apoptosis through Caspase pathway activation, which provides the reference for the clinical treatment ofhepatocellular carcinoma.

关 键 词：基因治疗 WNT/Β-CATENIN通路 溶瘤腺病毒 细胞凋亡 

分 类 号：R730.5[医药卫生—肿瘤]