论文部分内容阅读
ＰＦ４的Ｃ－末端１３肽和ＴＳＰ１的４２９～４５９片段３１肽通过Ｇｌｙ－Ｐｒｏ－Ｇｌｙ肽桥设计为融合蛋白ＴＳＦ．采用ｐＧＥＸ－２Ｔ载体在 Ｅ．ｃｏｌｉＪＭ１０９中获得了 ＴＳＦ蛋白的高效表达．采用 ＭＴＴ方法、损伤细胞迁移实验。鸡胚绒毛尿囊膜实验和小鼠移植肿瘤抑制实验，测定了ＴＳＦ及其相关物ＧＳＴ－ＴＳＦ，ＰＦ４（５８－７０），ＴＳＰ１（４２９～４５９）等对血管生成和肿瘤生长的抑制效应．结果显示ＴＳＦ特异抑制小牛主动脉血管内皮细胞的生长，并有明显的剂量依赖关系；非常显著抑制血管内皮细胞的运动迁移；显著抑制鸡胚绒毛尿囊膜的新生血管生成；上述抑制活性ＴＳＦ＞＞ＧＳＴ－ＴＳＦ＞ＰＦ４（５８～７０）＞ＴＳＰ１（４２９～４５９）．ＴＳＦ显著抑制小鼠Ｌｅｗｉｓ肺癌的生长，１．０μｍｏｌ／ｋｇ·ｄ的ＴＳＦ对Ｌｅｗｉｓ肺癌的抑瘤率达到６８．７５％．结果说明ＴＳＦ的重组设计是成功的，ＴＳＦ为一个人工重组的有效的血管生成抑制因子．
The C-terminal 13 peptide of PF4 and the 429-459 fragment 31 peptide of TSP1 were designed as fusion protein TSF by Gly-Pro-Gly peptide bridge. Using pGEX-2T vector in E. High expression of TSF protein was obtained in coliJM109. Using MTT method, damaged cell migration experiments. Inhibitory effects of TSF and its related substances GST-TSF, PF4 (58-70), TSP1 (429-459) on angiogenesis and tumor growth were measured in chick embryo chorioallantoic membrane experiments and mouse transplant tumor inhibition experiments. The results showed that TSF specifically inhibited the growth of calf aortic vascular endothelial cells in a dose-dependent manner; it significantly inhibited the migration of vascular endothelial cells; significantly inhibited the neovascularization of chicken embryo chorioallantoic membrane; the above inhibitory activity TSF >>GST-TSF>PF4(58-70)>TSP1(429-459). TSF significantly inhibited the growth of Lewis lung carcinoma in mice, and the tumor inhibition rate of Lewis lung carcinoma was 68.75% with 1.0μmol/kg·d TSF. The results showed that the recombinant design of TSF was successful, and TSF was an effective recombinant angiogenesis inhibitor.