应用endostatin蛋白治疗裸鼠SHG44脑胶质瘤的实验

Treatment of SHG44 gliomas in rat brain with endostatin protein

目的 克隆小鼠内皮抑素 (endostatin)基因 ,检测其表达蛋白生物学活性 ,应用该蛋白治疗裸鼠 SHG44脑胶质瘤 .方法 从小鼠胶原 c DNA用 PCR法克隆 endostatin基因 ,重组入 p UC19,测序后构建非融合表达载体 p DH- En-dostatin,使其在 DH5 α内经温度诱导表达 ,纯化该表达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 .经荷瘤裸鼠皮下注射该蛋白治疗其脑内胶质瘤 .结果 成功获得 endostatin基因 ,测序正确 .诱导表达蛋白 Mr为2 0× 10 3 ,具有抑制血管生成的活性 .该蛋白可抑制荷瘤裸鼠脑内胶质瘤生长 .结论 endostatin基因的表达和初步应用 ,为采用抗血管生成方法治疗脑胶质瘤等实体瘤研究奠定了实验基础 .

AIM To clone mouse endostatin gene, to detect the biological activities of its protein and to use it to treat SHG44 gliomas in the rat brain. METHODS Endostatin gene was cloned from the mouse collagen XVIII cDNA by PCR and recombined into the vector of pUC19. The gene was sequenced and non fusion expression vector pDH Endostatin was constructed. The recombinant mouse endostatin gene was expressed in DH5α at the condition of temperature induc tion. The protein activities were tested by Chicken Chorio Allantoic Membrane (CAM) assay and endothelial cell proliferation inhibitory assay. Endostatin was applied to the rat with tumor through hypodermic injection in order to cure the SHG44 gliomas of the rat brain. RESULTS The acquired endostatin gene sequence was correct and its protein size was 20×10 3. The protein possessed the anti angiogenesis activity and could inhibit the SHG44 gliomas growth of rat brain. CONCLUSION The expression and clinical application of mouse endostatin gene establish the foundation for the antiangiogenesis therapy of gliomas and other solid tumors.