http://www.100md.com 《第四军医大学学报》 2001年第23期
摘 要: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ⅩⅧcDNA用PCR法克隆endostatin基因，重组入pUC19，测序后构建非融合表达载体pDH-En-dostatin，使其在DH5α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20×103 ，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.
Treatment of SHG44gliomas in rat brain with endo┐statin protein
QU Yan，ZHANG Xiang，WU Jing-Wen，GAO Da-Kuan，GUO Yan，LI Xia
Institute of Neurosurgery of Chinese PLA，Xijing Hospital，Fourth Military Medical University，Xi'an710033，China
Abstract:AIM To clone mouse endostatin gene，to detect the biological activities of its protein and to use it to treat SHG44gliomas 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 DH5αat the condition of temperature induc-tion.The protein activities were tested by Chicken Chorio-Allantoic Membrane(CAM)assay and endothelial cell prolif-eration inhibitory assay.Endostatin was applied to the rat with tumor through hypodermic injection in order to cure the SHG44gliomas of the rat brain.RESULTS The acquired endostatin gene sequence was correct and its protein size was20×103 .The protein possessed the anti-angiogenesis activity and could inhibit the SHG44gliomas growth of rat brain.CONCLUSION The expression and clinical application of mouse endostatin gene establish the foundation for the an-tiangiogenesis therapy of gliomas and other solid tumors. ......
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