携带TAp63的双靶向溶瘤腺病毒抑制结肠癌干细胞生长  

Dual-regulated oncolytic adenoviruses carrying TAp63 gene inhibit the growth of colon cancer stem cells

作　　者：罗颀枫 杜敏[3] 代镇岭 付正伟 张振宇 Shiva Basnet 葛海燕 LUO Qifeng1,2, DU Min3, DAI Zhenling4, FU Zhengwei2, ZHANG Zhenyu2, Shiva BASNET2, GE Haiyan1,2(1. Department of General Surgery, Pinghu Second People＇ s Hospital, Pinghu 314200, China; 2. Department of General Surgery, Shanghai East Hospital, Tongji University, Shanghai 3. Department of Burn and Plastic Surgery, Qilu Hospital, Shandong University, Jinan 200120, China; 250012, China; 4. Department of General Surgery, Gongli Hospital, Pudong New Area, Shanghai 200135, Chin)

机构地区：[1]浙江省平湖市第二人民医院普外科,平湖314200 [2]同济大学附属上海市东方医院普外科,上海200120 [3]山东大学附属齐鲁医院烧伤整形科,济南250012 [4]上海市浦东新区公利医院普外科,上海200135

出　　处：《外科研究与新技术》2018年第2期77-82,共6页Surgical Research and New Technique

基　　金：浙江省医药卫生科技计划项目(2016KYB295)

摘　　要：目的构建携带TAp63的双靶向溶瘤腺病毒并检测其对结肠癌干细胞的抗瘤效应。方法应用流式细胞仪分选法筛选结肠癌干细胞;参照Adeasy系统构建重组腺病毒,并用重组腺病毒DNA作为模板,PCR反应后行琼脂糖凝胶电泳对病毒进行鉴定;应用CCK8和凋亡检测重组腺病毒对结肠癌干细胞和人正常肝细胞增殖及对结肠癌干细胞凋亡的影响。结果在HCT116细胞中,CD133/CD44双阳性细胞占(42.47±0.81)%(P<0.05);重组腺病毒Arsz-TAp63构建成功且无野生型污染,可抑制结肠癌干细胞HCT116^(CD133/CD44)增殖,且促进HCT116^(CD133/CD44)细胞凋亡,但对人正常肝细胞L-02增殖无影响。结论重组腺病毒Arsz-TAp63选择性抑制HCT116^(CD133/CD44)细胞增殖并诱导细胞凋亡。Objective To construct dual-regulated oncolytic adenoviruses carrying TAp63 gene and detect their antitumor effect on colon cancer stem cells. Methods Colon cancer stem cells were screened by flow cytometry. New recombinant adenoviruses were constructed according to the Adeasy system. The viral DNA acted as a template for PCR,then agarose gel electrophoresis assay was applied to confirm the viral DNA. CCK8 assay and cell apoptosis assay were used to detect the effects of recombinant adenoviruses on the proliferation of colon cancer stem cells and human normal liver cells,as well as on the apoptosis of colon cancer stem cells. Results The proportion of CD133/CD44 double positive cells was（ 42. 47 ± 0. 81） % in HCT116 cells（ P〈0. 05）. The recombinant adenoviruses Arsz-TAp63 were successfully constructed without wild-type adenovirus contamination,and were shown to inhibit the proliferation of colon cancer stem cells HCT116^（CD133/CD44）,but not human normal liver cells L-02,and promote the apoptosis of HCT116^（CD133/CD44） cells. Conclusion The recombinant adenoviruses Arsz-TAp63 selectively inhibit the proliferation and induce the apoptosis of HCT116^（CD133/CD44） cells.

关 键 词：结肠癌 干细胞 溶瘤腺病毒 

分 类 号：R735.3[医药卫生—肿瘤]