【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SMMC7721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αvcDNA克隆到腺病毒穿梭质粒pAd-Track巨细胞病毒(CMV)启动子下游,与腺病毒骨架质粒pAdEasy1在细菌内同源重组,在HEK293A细胞中包装成整合素αv重组腺病毒。以重组腺病毒感染SMMC7721细胞24h后,用蛋白质免疫印迹法(West-ernblot)检测整合素αv的表达;通过黏附实验研究其对SMMC7721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SMMC7721细胞后,整合素αv能够正确表达,并且SMMC7721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SMMC7721细胞的黏附中起重要作用,是肿瘤转移的重要分子基础。更多还原

【Abstract】 Objective To establish human integrinα_v adenovirus expressed in human liver cancer SMMC7721 cells and analyze the characteristics of integrinα_v adhesion to liver cancer cells. Methods Human integrinα_v cDNA was cloned into the plasmid pAdTrackCMV. Then, pAdTrackintegrinα_v 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 HEK293A cells to package and amplify recombinant adenovirus particles. Western blot showed that integrinα_v gene was exactly transcripted and expressed in SMMC7721 cells. Twenty-four hours after transfection, the effect of integrinα_v on adhesion of SMMC7721 cells was detected by adhesion experiment. Results Recombinant aenovirus vector of integrinα_v gene was successfully constructed and high titers of recombinant adnovirus was obtained . Adhesion cell count was significantly higher in integrinα_v-transfected cells than in untransfected and Ad-null-transfected cells (P<0.01). Conclusion Over-espression of integrinα_v promotes adhesion of SMMC7721 cells, is an important molecular mechanism of tumor metastasis.更多还原