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血管生成抑制素是新发现的一种血管生成抑制因子,能抑制肿瘤的生长和转移,有临床应用前景。根据血管生成抑制素是纤溶酶原的一个内片段的特点,以人纤溶酶原cDNA为模板,用PCR扩增出人血管生成抑制素基因,经序列分析后克隆至质粒pBV220转化大肠杆菌DH5α,在温度诱导下获得高效表达。SDS-PAGE及Western印迹分析显示:表达产物占菌体总蛋白的38％,相当于212mg/L,并具有免疫活性。生物活性分析结果表明,重组人血管生成抑制素能抑制bFGF诱导的CAM血管生成、抑制C57BL/6小鼠皮下原位B16黑色素瘤生长。
Angiostatin is a newly discovered angiogenesis inhibitor that inhibits the growth and metastasis of tumors and has clinical applications. According to the characteristic that angiostatin is an internal fragment of plasminogen, human angiostatin gene was amplified by PCR using human plasminogen cDNA as template, and cloned into plasmid pBV220 by sequence analysis to transform E. coli DH5α, under the temperature induced high expression. SDS-PAGE and Western blot analysis showed that the expressed product accounted for 38% of the total bacterial protein, equivalent to 212mg / L, and immunocompetent. The results of biological activity analysis showed that recombinant human angiostatin can inhibit bFGF-induced CAM angiogenesis and inhibit the growth of subcutaneous B16 melanoma in C57BL / 6 mice.