重组人内皮抑素的纯化及抗肿瘤活性研究

Study on Purification and Biological Activity of Recombinant Human Endostatin

[目的]从高效表达的基因工程菌中纯化重组人内皮抑素,对其抗肿瘤活性进行研究。[方法]经异丙基硫代鄄β鄄D鄄半乳糖苷(IPTG)诱导,重组人内皮抑素在大肠杆菌基因工程菌中以包涵体形式高效表达。通过凝胶层析纯化重组内皮抑素蛋白。应用鸡胚绒毛尿囊膜实验(CAM),肺癌细胞MTT试验及细胞迁移抑制实验,裸鼠皮下移植喉癌抑瘤实验、病理组织切片、免疫组化指标的测定等检测研究重组人内皮抑素的抗肿瘤活性。[结果]重组内皮抑素的复性率可达40%。重组内皮抑素在体外直接抑制肺癌细胞的增殖及迁移。用药21天,对裸鼠皮下移植喉癌的抑瘤率达到40.66%,HE染色、免疫组化及CAM实验表明其对肿瘤组织新生血管的生成有较强的抑制作用。[结论]重组人内皮抑素具有抑制肿瘤组织新生血管生成和直接抑制肿瘤细胞生长和迁移的双重抗肿瘤活性。

To purify recombinant human endostatin from high expression genetic engineering bacteria and analyze its anti-tumor activity.By isopropy1-beta-D-thiogalactopyranoside(IPTG) induction,recombinant human endostatin was expressed highly in Ecoli by inclusion.Recombinant human endostatin was purified by gel filtration chromatography.The biological anti-cancer activity of recombinant human endostatin were studied by chick embrgoallantoid membrane,MTT assay,cell migrate inhibition assay,the tumor growth of the laryngeal subcutaneously implanted in nude mice and pathological slice and immunohistochemistry.The restorable rate of recombinant human endostatin was 40%.Recombinant human endostatin could directly inhibit lung cell proliferation and migration.After 21 days of recombinant human endostatin administered,the tumor inhibition rate of the laryngeal subcutaneously implanted in nude mice was 40.66%.HE staining,immunohistochemistry,CAM identified recombinant human endostatin had a strong anti-angiogenesis activity on tumor tissue.[Conclusions]Recombinant human endostatin reveals effects to inhibite not only tumor tissue anti-angiogenesis but also tumor cell proliferation and migration.