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从鼠地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，并观察分泌上肖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。

Objective: To demonstrate that liposome mediated endostatin gene therapy is a viable approach for cancer antiangigensis therapy. Methods: Mouse endostatin cDNA was amplified from the murine liver by means of RT PCR. After being verified by sequence, secreted expression vector pSEC endo was constructed. The biological activity of the expressed endostatin was demonstrated by inhibition of proliferation of endothelial cells in response to stimulation by bFGF. The mechanism of this effect was studied by means of TDT dUTP nick end labeling assay and agarose electrophresis. By injection of liposomes complexed to murine endostatin cDNA, the inhibition of the growth of SMMC 7721 that implanted in nude mice was tested. Results: The sequence of mouse endostatin cDNA is identical with data reported from Olsen. The expressed endostatin could specifically inhibit the proliferation of endothelial cells in response to stimulation by basic fibroblast growth factor. The mechanism of this effect seems to be related with apoptosis. In addition, the efficacy of the endostatin vector treatment in reducing the volume of tumors implanted in nude mouse model was significant. Conclusion: These findings provide a basis for the further development of gene therapy with endostatin as an antiangiogenic gene.

国家 8 6 3项目 (86 3 Z2 0 0 1 0 4)