血管内皮生长因子部分多肽抗血管生成的研究

目的：观察血管内皮生长因子（VEGF）部分多肽（3－4外显子）抗血管生成的作用。方法：抽提LoVo细胞总RNA,进行RT－PCR,克隆VEGF部分多肽cDNA,构建VEGF部分多肽原核表达载体,用限制性酶切和DNA测序进行鉴定,通过聚丙烯酰胺凝胶电泳（SDS－PAGE）分析表达产物,表达产物经亲和层析纯化后,以人脐静脉内皮细胞（HUVEC）和鸡胚尿囊膜（CAM）血管测定其生物学活性。结果：表达产物以可溶性分子形成存在于菌体中,具有良好的抗原性和特异性,并具有抑制HUVEC增殖及CAM血管形成的活性。结论：VEGF部分多肽具有竞争抑制血管生成的功能,在肿瘤生物靶向治疗中具有一定潜在的价值。

Cloning,purification and biological activity of human vascular endothelial growth factor fragment in E. coli

OBJECTIVE: To observe the effect of human vascular endothelial growth factor (VEGF) fragment (3 approximately 4 exon) in E. coli on anti-angiogenesis. METHODS: Through RT-PCR amplification, endonuclease cut and DNA sequence analysis identification, hVEGF fragment cDNA was inserted into E. coli expression vector pTrcHis2A. The prokaryotic expression plasmid pTrcHis2A/VEGF(3 approximately 4) was constructed and transformed into TOP10F. RESULTS: After 8hr isopropy-beta-D-thiogalactoside (IPTG) induction, VEGF fragment was expressed in 15% of total proteins through sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The expressed protein was highly antigenic and specific. The VEGF fragment was further purified by affinity, which could inhibit HUVEC proliferation and neovascularization of the chick chorioallantoic membrane. CONCLUSION: VEGF fragment is anti-angiogenetic, which may potentially be used in oncologico-biological targeting therapy.