目的 :利用内皮细胞抑制素进行肿瘤抗血管基因治疗的尝试。方法 :用RT PCR从鼠肝脏扩增出内皮细胞抑制素cDNA ,经测序证实后 ,装入分泌表达载体pSEC hygromycin ,用DEAE 葡聚糖将其转染COS 7细胞 ,从mRNA水平及蛋白水平检测内皮细胞抑制素的表达 ,并观察分泌上清是否具有特异性抑制bFGF刺激的内皮细胞增殖作用 ,并通过TDT介导的dUTP缺口末端标记法和琼脂糖电泳来探讨其作用机制。在体外利用阳离子脂质体介导分泌表达载体pSEC endo进行抗肿瘤研究。结果 :测序结果表明 ,克隆的鼠内皮细胞抑制素cDNA与文献报道的完全一致 ,RT PCR显示转染后的COS 7细胞有内皮细胞抑制素mRNA表达 ,Western blot证实转染COS 7细胞上清及包浆内均有目的蛋白表达。其分泌上清能抑制bFGF刺激的内皮细胞增殖作用 ,且这种作用是通过诱导细胞凋亡实现的 ,动物模型显示所构建分泌表达的载体能在肿瘤内表达 ,有效抑制荷瘤裸鼠人肝癌细胞周围的微血管形成及肿瘤的生长。结论 :这些结果为抑血管基因在肝癌基因治疗中的进一步利用打下了基础 OBJECTIVE: An attempt to use anti-vascular gene therapy with endostatin. Methods: Endostatin cDNA was amplified from rat liver by RT PCR. After sequencing, it was inserted into the secretory expression vector pSEC hygromycin and transfected into COS 7 cells with DEAE dextran. The mRNA and protein levels Endostatin expression was observed, and whether the secretion supernatant had specific inhibitory effect on proliferation of endothelial cells stimulated by bFGF was investigated by TDT-mediated dUTP nick end labeling and agarose gel electrophoresis. Antitumor studies were performed in vitro using cationic liposome-mediated secretion expression vector pSEC endo. Results: The sequencing results showed that the cloned rat endostatin cDNA was completely identical with that reported in the literature. RT-PCR showed that the transfected COS 7 cells had the expression of endostatin mRNA. Western blot confirmed that the transfected COS 7 cells supernatant and Pulmonary plasma protein expression. The secreted supernatant can inhibit bFGF-stimulated endothelial cell proliferation, and this effect is achieved through the induction of apoptosis, animal models show that the expression of the constructed expression vector can be expressed in the tumor, effectively inhibiting human hepatoma in nude mice Microvascular formation around cells and tumor growth. Conclusions: These results provide a basis for the further utilization of anti-angiogenic genes in HCC gene therapy