人纤溶酶原K1-3基因的表达及抗肿瘤作用研究

Expression of kringle 1-3 domain fragment of human plasmihogen in E.coli and the anti-tumor activity of its product

目的:研究人纤溶酶原K1-3基因的原核表达及其产物K1-3蛋白的抗肿瘤活性。方法:构建含K1-3基因的表达载体pBV-K13;转化E.coliDH5α;热诱导表达菌株E.coliDH5α(pBV-K13)的K1-3基因表达;表达产物经溶解、复性和纯化后进行鸡胚绒毛尿囊膜(Chorioallantoic membrane,CAM)活性分析和小鼠B16黑色素瘤抑制实验。结果:SDS-PAGE和Western blot分析结果证实K1-3基因在大肠杆菌中获得高效表达,表达产物对人纤溶酶原抗血清具有特异免疫活性,并抑制CAM毛细血管生成及小鼠B16黑色素瘤生长。结论:原核表达的K1-3蛋白具有抑制血管生成和抗肿瘤作用。

Objective： To study the expression of kringle 1-3 domain fragment of human plasmihogen in E.coil and the anti-tumor activity of its product.Methods：The K1-3 domain fragment was cloned in expression vector pBV220,the resulted recombinant plasmid pBV-K13 was transformed into E.coli DH5a and its product was purified and assayed its bioaetivity. Results： K1-3 domain fragment was expressed in E. coli DH5α. The results showed the expressed product covered 20% of the total bacterial protein on SDS-PAGE and the Western blot analysis showed that the product had immunological specificity with the antiserum of human plasminogen and inhibits the growth of chorioallantoic membrane（CAM） angiogenesis and mouse B16 melanoma. Conclusion： Human plasminogen K1-3 domain fragment was expressed in E. coli; the expressed product has anti-angiogenesis and antitumor activity.