重组人p43蛋白抗血管生成活性及其机制

Anti-angiogenetic effect of recombinant human p43 and its mechanism

为了研究重组人p43(YS-1)蛋白对血管生成的影响及其机制,并考察其对肺癌实体瘤的抑制作用。采用内皮细胞增殖实验、细胞迁移实验、管腔形成实验、大鼠动脉环体外实验、鸡胚尿囊膜体内实验考察YS-1的抗血管生成作用;采用Western印迹法检测细胞内磷酸化MEK1/2、Akt蛋白质的表达;选用人肺腺癌A-549裸小鼠移植瘤模型检测YS-1的体内抗肿瘤活性。结果发现,YS-1能明显抑制内皮细胞迁移和管腔形成,可以抑制体外培养的大鼠动脉环微血管样结构及鸡胚尿囊膜血管,YS-1可以明显抑制血管内皮生长因子(VEGF)诱导的内皮细胞中VEGFR2及下游MEK1/2及Akt的磷酸化,YS-1高剂量(10 mg/kg)能显著抑制肺癌A549移植瘤的生长。研究结果表明:YS-1能抑制人肺腺癌A-549裸鼠移植瘤的生长,其作用机制可能跟抑制VEGF引起的VEGFR2活化及信号转导而影响血管生成有关。

The purpose of this study was to investigate the effects and mechanism of recombinant human p43 （YS-1） on angiogenesis, and explore the antitumor activity of YS-1 in nude mice xenografted with lung adenocar- cinoma A-549. Endothelial cell proliferation assay, cell migration assay, tube formation assay, rat aortic ring assay and chicken chorioallantoic membrane（CAM） assay were used to evaluate the anti-angiogenetic activity of YS-1, and Western blot analysis was used to investigate the related mechanism. A model of lung adenocarcinoma A-549 xenograft in nude mice was used to explore the antitumor efficacy of YS-1. It was found that YS-1 inhibited endo- thelial cell migration and tube formation, and YS-1 obviously inhibited microvessel sprouting from rat aortic rings and the vessel growth in CAM. In addition, YS-1 attenuated the activation of VEGFR2, MEK1/2 and Akt in response to vascular endothelial growth factor （VEGF）. Moreover, high dose of YS-1 （ 10 mg/kg） was shown to exert a remarkable antitumor effect in the examined model. Therefore, YS-1 can inhibit adenocarcinoma A-549 xenograft in nude mice and its mechanism may be related to the inhibition of angiogenesis resulting from the attenuation of activated VEGFR2 and downstream signaling molecules.