重组腺病毒介导反义c-myc基因对人肝癌细胞系的治疗作用

Adenovirus mediated antisense c-myc fragment gene therapy for human liver cancer cells in vitro and in vivo

目的：探讨重组腺病毒介导反义 c-myc基因(Ad-ASmyc)治疗人肝癌细胞的作用。方法：观察 Ad-ASmyc对人肝癌细胞系的转导效率，通过细胞生长曲线、克隆形成实验、 DNA片段化分析、RT-PCR、裸鼠皮下移植瘤治疗实验，分析 Ad-ASmyc对人肝癌细胞系 Bel-7402、 QSG-7701、 SMMC-7721和 HCC-9204细胞生长和 c-myc基因表达及裸鼠肿瘤生长的抑制作用。结果： Ad-ASmyc可高效转导人肝癌细胞系，抑制细胞生长；转染细胞克隆形成能力降低，克隆成活率为对照组的53.9％～69.1％，c-myc基因表达下降；Ad-ASmyc处理肝癌细胞， DNA凝胶电泳出现明显的梯形条带，瘤内注射 Ad-ASmyc可抑制裸鼠皮下移植瘤生长。 结论：重组腺病毒介导的反义 c-myc基因转移，有可能成为肝癌基因治疗的一种有效方法。

Objective: The current study was designed to investigate the potential of antisense c-myc gene therapy for hepatocellular carcinoma. Methods:The responses of human hepatocellular carcinoma cell lines,Bel-7402, QSG-7701, SMMC-7721 and HCC-9204 to recombinant adenovirus encoding antisense c-myc RNA fragment (Ad-ASmyc)were analyzed in vitro. Meanwhile,effects of Ad-ASmyc on Bel-7402 subcutaneous tumors in nude mice were studied in vivo. Results: Growth inhibition and reduction of the ability of colony formation were observed in vitro in all four hepatocellular carcinoma cell lines with Ad-ASmyc.Obvious apoptosis was observed in the infected tumor cells as shown by morphological changes under microscope and DNA fragmentation. In vivo studies,it was showed that the growth of subcutaneous tumors of Bel-7402 cells was significantly inhibited by intratumor injection of Ad-ASmyc. Conclusion:Adenovirus mediated antisense-myc is a potential clinical utility in gene therapy for hepatocellular carcinoma.