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目的 克隆小鼠 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 从小鼠胶原 c DNA用 PCR法克隆 endostatin基因 ,重组入p U C19,测序后构建非融合表达载体 p DH- endostatin,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 .经荷瘤大鼠皮下注射该蛋白治疗其脑胶质瘤 .结果 成功获得 5 5 1bp的endostatin基因 ,经测序正确 .该诱导表达蛋白 Mr2 0 0 0 0 ,具有抗血管生成活性 .应用该蛋白 10或 2 0 mg· kg- 1· d- 1可抑制荷瘤大鼠脑胶质瘤血管生成和肿瘤生长 .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细胞瘤的研究奠定了实验理论基础
Objective To clone the endostatin gene of mouse and detect the biological activity of its expressed protein, and use this protein to treat rat C6 glioma.Methods The endostatin gene was cloned from mouse collagen c DNA by PCR and cloned into pUC19 and sequenced The non-fusion expression vector p DH-endostatin was induced to express in DH5α by temperature, the expressed protein was purified and the activity was detected by chick chorioallantoic membrane assay and endothelial cell inhibition assay. Its glioma.Results The endostatin gene of 5 5 1bp was successfully obtained and sequenced correctly.This protein was induced to have anti-angiogenic activity Mr 2 000. The protein was applied at 10 or 20 mg · kg -1 · d - 1 can inhibit glioma angiogenesis and tumor growth in tumor - bearing rats.Conclusion The expression of Endostatin gene and its preliminary application lay the theoretical foundation for the study of anti - angiogenesis in the treatment of glioma