HSV-TK/GCV系统杀伤肝癌细胞的在体研究

GENE THERAPY FOR HEPATOCELLULAR CARCINOMA WITH HSV TK/GCV SYSTEM IN VIVO

目的和方法：将可产生含有ＨＳＶＴＫ基因逆转录病毒的包装细胞与ＢＥＬ７４０２肝癌细胞混合后，接种于裸鼠皮下，复制肝癌模型。确定ＨＳＶＴＫ基因转移后，给予动物注射ＧＣＶ，观察ＨＳＶＴＫ／ＧＣＶ系统基因治疗实验性肝癌的在体疗效。结果：在体条件下，逆转录病毒可将ＨＳＶＴＫ基因部分转导至肝癌细胞；ＧＣＶ作用后，ＴＫ＋中瘤细胞的生长受到明显抑制；电镜观察发现，ＧＣＶ杀伤ＴＫ＋肿瘤细胞的作用主要体现在细胞核；虽然组化染色分析表明ＴＫ基因转移效率只有１０％～３０％，但测量肿瘤体积显示ＨＳＶＴＫ／ＧＣＶ系统杀伤肝癌细胞的效果依然明显，故其作用机制可与“旁观者效应”有关。结论：ＨＳＶＴＫ／ＧＣＶ“自杀”

Aim and Methods: We established the hepatic tumor models in nude mice (BALB/c) by hypodermic inoculation of hepatocellular carcinoma cell line BEL 7402 and packaging cell line PA317(tk +/tk -). After confirmation of tk gene transfer, GCV treatment was initiated to investigate the effect of HSV tk/GCV system in vivo. Results:Thee HSV tk gene could be efficiently transferred into hepatoma cells via retrovirus in vivo. The growth of tk + tumors was remarkably inhibited by GCV. Electron microscopy revealed the most significant changes in tk + cells appeared in the nuclei. Although the histochemical examination demonstrated that the efficiency of the gene transfer was about 10%～30%, the killing effect of HSV tk/GCV system on hepatocellular carcinoma was significant and was probably associated with 'bystander effects'. Conclusion: These results suggested that HSV tk/GCV system might provide a potential approach for future clinical application.