【摘要】 目的通过比较野生及重组人类酸性成纤维细胞生长因子(waFGF,raFGF)的蛋白结构稳定性、细胞增殖活性的影响,探讨核转位序列的敲除对aFGF促分裂活性的影响。方法Western-blot检测waFGF和raFGF两种蛋白结构稳定性;MTT法检测NIH3T3细胞增殖,即促细胞分裂活性;流式细胞术检测其对细胞周期的影响;同时,检验肝素(HS)对waFGF和raFGF有关生物学作用。结果waFGF的结构稳定性大于raFGF;细胞增殖实验表明,waFGF的促分裂活性大于raFGF,P<0.05。在肝素的参与下,waFGF和raFGF表现出更大的促分裂活性;结论与waFGF比,raFGF的结构稳定性小于waFGF,具有较弱的促分裂性,仍具有刺激DNA合成和细胞增殖的能力。肝素具有增强aFGF活性的作用,可作为aFGF体外实验中模仿机体内环境的辅助因子。更多还原

【Abstract】 Objective To study the effect of aFGF with the nuclear translocation sequence knocked out on the mitogenic activity.Methods Western bolt for the assays of the structural stability of both this two proteins.MTT for the cell proliferation assays.The role of heparin in aFGF mitogenic activity was also tested in this study.Results For the mitogenic activity,there were significant differences between wFGF and raFGF(P<0.05).Compared to waFGF,the raFGF showed the weaker activity.Under the treatment of test compounds,With the presence of heparin,either waFGF or raFGF showed bigger activity than that without the presence of heparin.Conclusion The mitogenic activity of raFGF is weaker than that of waFGF.They all can stimulate DNA synthesis and cell proliferation.The structural stability of raFGF is lowered than that of waFGF.Heparin can enhance the aFGF activity,and can act as a asistantfactor to imitate the the internal environment in aFGF in vitra test.更多还原