摘要
目的:研究微管抑制剂C9对新生血管生成的抑制作用及其机制。方法:利用流式细胞术和TUNEL法,检测C9逆转血管内皮生长因子(vascular endothelial growth factor,VEGF)和碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)抗无血清诱导的内皮细胞凋亡;Western Blot法检测C9对生长因子VEGF和bFGF刺激的下游与新生血管生成相关的两条通路Raf-Mek-Erk和PI3K-AKT中Raf,Mek,Erk,Akt磷酸化影响。结果:C9能够逆转VEGF和bFGF抗无血清诱导的内皮细胞凋亡,可以剂量依赖性地下调生长因子VEGF和bFGF下游与新生血管生成相关的两条通路Raf-Mek-Erk和PI3K-AKT中Raf,Mek,Erk,Akt磷酸化,但是对VEGF受体KDR及bFGF受体FGFR1的磷酸化没有影响。结论:C9主要通过直接抑制Raf-Mek-Erk信号通路而抑制生长因子的抗内皮细胞凋亡作用,并产生抑制新生血管生成作用。
Objective： To investigate mechanism underlying the anti-angiogenetic effect of C9, a new tubu- lin inhibitor. Methods： Antagonizing effect of C9 on anti-apoptotic activity of VEGF and bFGF was determined by FACS and TUNEL assay in endothelial cells cultured in serum-free media. Phosphorylation of c-Raf, Mek, Erk and Akt was detected by Western blot after treatment with various concentrations of C9. Results： FACS and TUNEL assay showed that C9 antagonized the anti-apoptosis activity of VEGF and bFGF on apoptosis induced by serum-free culture in endothelial cells. Western Blot revealed that C9 at various concentrations downregulated phosphorylation of c-Raf, Mek, Erk and Akt, but did not affect the phosphorylation of KDR or FGFR1. Conclusion： C9 inhibits the angiogenesis process by targeting the Raf-Mek-Erk-MAPK pathway.
出处
《中国新药杂志》
CAS
CSCD
北大核心
2011年第17期1703-1710,共8页
Chinese Journal of New Drugs
基金
国家自然科学基金(30572201)
