复制缺陷型腺病毒PCA的构建及其对结肠癌细胞生长的影响  

Establishment of Replication-deficient Recombinant Adenovirus PCA and Its Influence on Cell Growth of Colon Cancer

作　　者：黄越[1] 陈文铎[1] 周炜[1] 董淼[1] 高永[1] 许韶威 徐涛[1] 唐斌[1] 

机构地区：[1]浙江理工大学生命科学学院,杭州310018

出　　处：《浙江理工大学学报（自然科学版）》2013年第6期901-905,918,共6页Journal of Zhejiang Sci-Tech University(Natural Sciences)

基　　金：国家自然基金资助项目(30721712)

摘　　要：通过同源重组方式,利用Adeasy系统成功构建携带p14ARF基因的复制缺陷型腺病毒,命名为AdpShuttle-CMV-p14ARF(PCA),并研究其对结肠癌细胞株SW480及HT-29的杀伤作用。PCR方法鉴定阳性重组子病毒;MTT法检测PCA对细胞增殖的影响;Hoechst33258荧光染色法检测细胞凋亡形态学变化;结晶紫染色法定性分析PCA对两种结肠癌细胞的杀伤作用。结果显示:经目的病毒PCA作用,SW480及HT-29出现明显病变和生长抑制现象,并呈现出时间和浓度依赖性;Hoechst染色后观察到细胞内出现凋亡小体,并且数目随药物处理浓度的升高而增多。This paper successfully establishes replication-deficient recombinant adenovirus called AdpShuttle-CMV-p14ARF （PCA） which carries p14ARF gene using Adeasy system through homologous recombination; studies its lethal effect on colon cancer cell strains SW480 and HT-29; identifies positive recombinant subvirus with PCR method; detects the influence of PCA on cell proliferation with MTT method; detects morphological change of cell apoptosis with Hoechst33258 fluorescent staining method; and conducts qualitative analysis on the lethal effect of PCA on two colon cancer cells with crystal violet staining.The result shows that SW480 and HT-29 have obvious lesion and growth inhibition phenomena and present time and concentration dependence under the action of target virus PCA; apoptotic body occurs in cells after Hoechst staining and its number increases with the increase of medication concentration.

关 键 词：Ad-p14-ARF 结肠癌 MTT 结晶紫染色 细胞凋亡 

分 类 号：Q789[生物学—分子生物学]