BTG2基因重组慢病毒载体的构建及其在肺癌A549细胞中的表达与功能  

Construction of Recombinant Lentiviral Vector pCDH-BTG2 and Its Expression and Function in A549 Cells

作　　者：生秀梅[1] 王正新[1] 

机构地区：[1]江苏大学医学院生物化学教研室,镇江212013

出　　处：《福建医科大学学报》2016年第2期74-77,共4页Journal of Fujian Medical University

基　　金：国家自然科学基金(31000046);国家博士后基金面上项目(2015M571702);江苏大学高级人才启动基金(11JDG063)

摘　　要：目的构建重组慢病毒载体pCDH-BTG2,并检测其转染肺癌A549细胞后的表达与功能。方法通过PCR技术以含B细胞转位基因(BTG2)的cDNA克隆质粒(HsCD00330081)为模板获得BTG2基因片段,将此片段克隆至慢病毒载体pCDH-CMV-MCS-EF1-Puro,形成pCDH-BTG2,转染PC3细胞检测BTG2的表达;将重组慢病毒载体质粒pCDH-BTG2包装成重组慢病毒颗粒,感染肺癌A549细胞后,通过蛋白免疫印迹法检测BTG2的表达,细胞计数法检测BTG2对A549细胞生长的影响。结果 pCDH-BTG2重组质粒构建成功,转染PC3细胞后可检测BTG2蛋白的表达,pCDH-BTG2包装成慢病毒感染A549细胞后,显著抑制A549细胞的生长。结论成功构建了重组慢病毒载体pCDH-BTG2,并发现BTG2抑制A549细胞的生长,为研究BTG2的功能奠定了基础,同时为肺癌的药物研究提供了靶点。Objective To construct recombinant lentiviral vector pCDH-BTG2 and detect its expression and function in A549 cells. Methods cDNA clone plasmid（HsCD00330081）was used as template to amplify the fragment BTG2 by PCR. The fragment was cloned into pCDH-CMV-MCS-EF1-Puro to generate recombinant lentiviral vector pCDH-BTG2. PC3 cells were transfected with pCDH-BTG2 and its expression was detected by Western-blot. Recombinant lentivirus was produced by 293 Tcells following co-transfection of pCDH-BTG2 with the packaging plasmids. The resulting recombinant lentivirus Lenti-BTG2 was then used to infect A549 cells. BTG2 expression was detected by Western-blot analysis,and cell growth assays were performed to check the effect of BTG2 on A549cells. Results The recombinant lentiviral vector pCDH-BTG2 was successfully constructed and could be detected by Western-blot after transfecting PC3 cell. Lentivirus Lenti-BTG2 was successfully constructed and could infect A549 cells. Expression of BTG2 obviously inhibited cell growth of A549 cells. Conclusion The lentiviral vector pCDH-BTG2 was successfully constructed,which lays the foundation for the research of BTG2 function. We also found that BTG2 inhibits lung cancer cell growth,which provides a potential therapeutic agent for lung cancer.

关 键 词：慢病毒属 基因 基因表达 肺肿瘤 肿瘤细胞 培养的 重组 遗传 

分 类 号：R734.2[医药卫生—肿瘤]