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两元腺病毒载体系统转移bax基因诱导肝癌细胞凋亡的研究

Study on the bax Gene Transferred by a Binary Adenoviral Vector system to Induce Apoptosis of Human Hepatocellular Carcinoma Cells
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摘要 目的 采用两元腺病毒载体系统,介导bax基因转移至培养的肝癌细胞SMMC-7721,探讨腺病毒介导转移bax基因抗肝癌的活性。方法 常规方法在293细胞中扩增腺病毒Ad/GT-Bax和Ad/PGK-GV16,CsCl密度梯度离心法纯化病毒后测定病毒滴度。结果 (1)Bax蛋白表达情况:感染Ad/GT-Bax和Ad/PGK-GV16的SMMC-7721细胞,其在24、48和72小时的Bax蛋白表达量依次为阴性对照细胞的2.5、3.1和3.9倍;(2)PI染色后,感染Ad/GT-Bax和Ad/PGK-GV16的SMMC-7721细胞出现亚二倍体峰,该峰面积占总面积的值在24、48、72小时时依次为31.65%、50.44%和61.81%。结论 两元腺病毒载体系统介导转移Bax基因可诱导人肝癌SMMC-7721细胞凋亡,可望成为治疗肝癌的有力工具。 Objective To investigate the anti-hepatocellular carcinoma activity of bax gene transferred to the hepatocellular carcinoma cell line SMMC-7721 by a binary adenoviral vector system. Methods The adenoviral vectors, Ad/GT-Bax and Ad/PGK-GV16, were amplified in cell line 293 conventionally and purified by CsCl density gradient centrifugation, and then the liters were detected. The rate of cell apoptosis was detected by PI dyeing after the human hepatocellular carcinoma cells were infected with MOI 200: 1. and the bax protein expressed by cells was detected by immunoblot assay. Results ( 1) The expression qualities of bax protein of the SMMC-7721 cells infected by Ad/GT-Bax and Ad/PGK-GV16 were 2.5, 3.1 and 3.9 times to the negative controll cells after 24, 48 and 72 hours respectively, (2) Hypodiploidy peak appeared in the SMMC-7721 cells infected by Ad/GT-Bax and Ad/PGK-GV16 after dyed with PI. The percents of the peak area in total area were 31.65% , 50.44% and 61.81% after 24, 48 and 72 hours, respectively. Conclusion Bax gene transferred by the binary adenovira] vector system induce the human hepatocellular carcinoma cell line SMMC-7721, which might be a potential drug for hepatocellular carcinoma cell.
出处 《浙江预防医学》 2003年第3期5-6,9,共3页 Zhejiang Journal of Preventive Medicine
关键词 两元腺病毒载体系统 BAX基因 肝癌 细胞凋亡 Hepatocellular carcinoma Cell apoptosis bax gene
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