aFGF核转位序列的敲除后蛋白结构稳定性及促分裂活性的变化  

The effect of the nuclear translocation sequence-knocked out on the mitogenic activity of aFGF

作　　者：吕文天[1] 付士波[1] 彭菲[2] 吕喆[1] 顾加祥[3] 王志成[1] 李校堃[4] 龚守良[1] 

机构地区：[1]吉林大学公共卫生学院卫生部放射生物学重点实验室,吉林长春130021 [2]暨南大学药学院生物工程教研室,广州510632 [3]吉林大学中日联谊医院手外科,长春130033 [4]温州医学院生物与天然药物研究院,温州325035

出　　处：《中国实验诊断学》2006年第10期1122-1125,共4页Chinese Journal of Laboratory Diagnosis

基　　金：国家863专题项目(2002AA2Z3318)

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。Objective To study the effect of aFGF wih the nuclear transloeation 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 tiffs 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 flint 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 asistant factor to imitate the the internal environment in aFGF in vitra test.

关 键 词：重组酸性成纤维细胞生长因子 促分裂性 结构稳定性 肝素 细胞增殖 

分 类 号：R346[医药卫生—基础医学]