Cyclin B1正、反义全长cDNA腺病毒载体的构建及对HeLa细胞增殖和凋亡的影响  

Construction of Recombinant Adenovirus Containing Full-length Sense and Antisense Cyclin B1 cDNA and Their Impact on Proliferation and Apoptosis of HeLa Cells

作　　者：李洁[1,2] 陈平[2] 胡火珍[1,2] 

机构地区：[1]四川大学生命科学学院生物资源与生态环境教育部重点实验室,成都610064 [2]四川大学华西医院生物治疗国家重点实验室

出　　处：《四川大学学报（医学版）》2009年第5期765-769,共5页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(批准号30672204)资助

摘　　要：目的利用pAd/CMV/V5-DEST腺病毒载体系统构建含有人细胞周期蛋白B1(Cyclin B1)正、反义全长cDNA的重组腺病毒,并研究其对人宫颈癌细胞株HeLa增殖及凋亡的影响。方法通过RT-PCR获取Cyclin B1全长cDNA,分别以正、反方向插入pENTR11,与pAd/CMV/V5-DEST进行同源重组后获得正确的重组腺病毒质粒,经293A细胞包装扩增获得重组病毒颗粒。重组腺病毒体外感染HeLa细胞,通过细胞计数和流式细胞术观测其对细胞增殖及凋亡的影响。结果Cyclin B1基因成功克隆到载体上,并经293A细胞包装出病毒颗粒。此重组腺病毒感染HeLa细胞后,反义Cyclin B1能明显抑制细胞的生长并且促进细胞凋亡。结论成功构建了携带人Cyclin B1正、反义全长cDNA的重组腺病毒载体,此载体可在HeLa细胞中发挥生物学作用。Objective To construct a recombinant adenovirus vector carrying the full-length sense and antisense cyclin B1 cDNA of human, and to investigate the influence of the recombinant adenoviruses on the cultured human cervical carcinoma cell line HeLa. Methods The recombinant adenovirus vectors were constructed by homologous recombination technique. The recombinant adenoviruses were packaged and amplified in 293A cells. Then the recombinant adenoviruses were used to infect HeLa cells. The influence was determined by cell counting and flow cytometry. Results The HeLa cells infected with antisense recombinant adenovirus exhibited significant cell apoptosis and growth inhibition compared to the normal HeLa cells and the cells infected with sense recombinant adenovirus. Conclusion Recombinant adenoviruses vectors containing the full-length sense and antisense eDNA of human cyclin B1 is successfully constructed, which has laid a foundation for further studies on its anticancer functions.

关 键 词：细胞周期蛋白B1 重组腺病毒 基因治疗 肿瘤 

分 类 号：R737.33[医药卫生—肿瘤]