李旭艳 刘靖华 李志杰 莫永炎 刘亚伟 刘志锋 邓鹏 姜勇

    【摘要】 目的 构建表达人整合素αv亚基(整合素αv)重组腺病毒，并在人肝癌细胞株SMMC-7721细胞中表达，探讨整合素αv对肝癌细胞黏附特性的影响。方法 将人整合素αv cDNA克隆到腺病毒穿梭质粒pAdTrack-巨细胞病毒(CMV)启动子下游，与腺病毒骨架质粒pAdEasy-1在细菌内同源重组，在HEK-293A细胞中包装成整合素αv重组腺病毒。以重组腺病毒感染SMMC-7721细胞24 h后，用蛋白质免疫印迹法(Western blot)检测整合素αv的表达；通过黏附实验研究其对SMMC-7721细胞黏附的影响。结果 构建了整合素αv重组腺病毒载体并制备出高滴度重组腺病毒。重组腺病毒感染SMMC-7721细胞后，整合素αv能够正确表达，并且SMMC-7721细胞黏附率显著高于对照组和空白组(P<0.01)。结论 整合素αv基因在SMMC-7721细胞的黏附中起重要作用，是肿瘤转移的重要分子基础。

    【关键词】 整合素； 细胞黏附； 腺病毒载体

    Construction of a recombinant adenoviral vector carrying integrin-αv and the influence of integrin-αv on the adhesion characteristics of SMMC-7721 cells LI Xu-yan, LIU Jing-hua, LI Zhi-jie, MO Yong-yan, LIU Ya-wei, LIU Zhi-feng, DENG Peng, JIANG Yong. Department of Pathophysiology, Key Laboratory of Functional Proteomics of Guangdong Province, Southern Medical University, Guangzhou 510515,Guangdong, China Corresponding author:JIANG Yong (Email:yjiang@fimmu.com)

    OBJECTIVE: To establish human integrin-αv adenovirus expressed in human liver cancer SMMC-7721 cells and analyze the characteristics of integrin-αv adhesion to liver cancer cells. METHODS: Human integrin-αv cDNA was cloned into the plasmid pAdTrack-CMV. Then, pAdTrack-integrin-αv was cotransformed with adenoviral backbone vector into E. coli strain BJ5183. The recombinant adenoviral plasmid was identified by restriction analysis with Pac I and transfected into HEK-293A cells to package and amplify recombinant adenovirus particles. Western blot showed that integrin-αv gene was exactly transcripted and expressed in SMMC-7721 cells. Twenty four hours after transfection, the effect of integrin-αv on adhesion of SMMC-7721 cells was detected by adhesion experiment. RESULTS: Recombinant aenovirus vector of integrin-αv gene was successfully constructed and high titers of recombinant adnovirus was obtained. Adhesion cell count was significantly higher in integrin-αv transfected cells than in untransfected and Ad null transfected cells (P<0.01). CONCLUSION: Over espression of integrin-αv promotes adhesion of SMMC-7721 cells, is an important molecular mechanism of tumor metastasis. ......