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目的 观察腺病毒介导的单纯疱疹病毒胸苷激酶基因 (HSV tk) /丙氧鸟苷 (GCV )自杀基因系统在体内外对人肝癌细胞的杀伤效应。方法 在体外按MOI值为 10 0、10、1、0用重组腺病毒转染人肝癌细胞BEL 740 2和SMMC 772 1,48h后用噻唑蓝 (MTT)法测定细胞存活率。在两种肝癌裸鼠模型上 ,瘤体内注射重组腺病毒 ( 1× 10 12 pfu/L) 0 .1ml,GCV作用后观察抑瘤效果。 结果 体外MOI为 10 0时 ,可杀死 99.8%的BEL 740 2细胞和 17.8%的SMMC 772 1细胞 ,两者差异有显著性 ( P <0 .0 5 )。体内BEL 740 2组肿瘤生长明显受到抑制。结论 在体内外 ,腺病毒介导的含AFP调控序列的HSV tk/GCV自杀基因系统对甲胎蛋白 (AFP)阳性肝癌细胞具有特异性杀伤作用 ,该系统可望用于肝癌的特异性基因治疗。
Objective To observe the killing effect of adenovirus-mediated HSV tk/GCV suicide gene system on human hepatoma cells in vitro and in vivo. METHODS: Cell viability was measured by MTT assay after in vitro transfection of human hepatoma cells BEL 740 2 and SMMC 772 with MOI values of 10 0, 10, 1, and 0 for 48 h. In two nude mice models of hepatocellular carcinoma, 0.1 ml of recombinant adenovirus (1×10 12 pfu/L) was injected intratumorally, and the anti-tumor effect was observed after GCV treatment. Results In vitro MOI at 10 0 killed 99.8% of BEL 740 2 cells and 17.8% of SMMC 772 1 cells, with a significant difference (P < 0.05). In vivo BEL 740 2 tumor growth was significantly inhibited. Conclusion Adenovirus-mediated HSV tk/GCV suicide gene system containing AFP regulatory sequences has specific killing effect on alpha-fetoprotein (AFP)-positive hepatocellular carcinoma cells. This system is expected to be used for the specific gene therapy of liver cancer. .