小鼠内皮抑素基因的克隆、表达及其抑瘤效应研究

克隆小鼠内皮抑素（ｅｎｄｏｓｔａｔｉｎ）基因，检测其表达蛋白的生物学活性，应用该蛋白治疗大鼠Ｃ６脑胶质瘤。方法：采用ＲＴ－ＰＣＲ法，从小鼠肝组织克隆ｅｎｄｏｓｔａｔｉｎ基因，重组入ｐＵＣ１９，测序后构建非融合表达载体ｐＢＶ２２０－ｅｎｄｏｓｔａｔｉｎ，使其在ＤＨ５α内经温度诱导表达，纯经ｅｎｄｏｓｔａｔｉｎ蛋白并用鸡胚绒毛膜尿囊膜实验和内皮细胞抑制实验检测其活性，经荷Ｃ６胶质瘤大鼠皮下注射该蛋白，

The research of clone and expression of mouse endostatin gene and detection its inhibition effect to the C6 glioma

Objective: To clone mouse endostatin gene, detect its biological activities and use its protein to treat C6 gliomas of the rat brain. Methods: Endostatin gene was cloned from the mouse liver by RT-PCR and recombined into the vector of pUC19. The gene was sequenced and non-fusion expression vector pBV220-Endostatin was constructed. The recombinant mouse endostatin gene was expressed in DH5a at tempendure induction. The Protein activities were tested by chicken chorio-allantoic membrane (CAM) assay and endothelial cell proliferaion inhibitory assay. Through hypodermic injection, endostatin was applied to the rat bearing C6 glioma in order to determine its inhibition effect to the tumor.Results: The endostatin gene sequence obtained was cormct and its protein size was 20 kDa. The protein appeared the anti-angiogenesis artivity and inhibited the C6 glioma prowht. Conclusions: The successful clone, expression and application of mouse endostatin gene lay the foundation for the aniangiogenesis therapy of the solid tumors.