牡蛎多肽抗血管生成作用研究

Antiangiogenic Effect of Oyster Polypeptide (OPP)

背景与目的血管发生、新血管的形成是肿瘤生长及其转移灶形成的根本原因,因此,抗血管生成治疗已成为目前一种极具潜能的肿瘤治疗策略。本实验利用鸡胚胎模型和人脐静脉内皮细胞(Human umbilical vein en-dothelial cells,HUVECs)对牡蛎多肽(Oyster polypeptide,OPP)在体内及体外的抗血管生成作用及其机制进行了研究。方法利用鸡胚胎绒毛尿囊膜血管发育研究体内OPP对血管形成作用;MTT、平板划痕、Transwell板、管腔形成及电镜等方法观察OPP对HUVECs的增殖、迁移、侵袭等作用。结果体内实验表明OPP能明显抑制鸡胚胎绒毛尿囊膜血管的形成。MTT结果表明OPP能明显抑制HUVECs的增殖,IC50为400μg/mL。平板划痕法结果表明,200μg/mL、400μg/mL和800μg/mL OPP作用12h对HUVECs迁移的抑制率分别为18.75%、37.93%和74.07%;侵蚀实验结果显示200μg/mL、400μg/mL和800μg/mL的OPP对HUVECs迁移的抑制率分别为15.5%、37.29%和67.24%(P<0.05)。管腔形成结果表明200μg/mL、400μg/mL和800μg/mL的OPP对HUVECs(作用10h)小管形成的抑制率分别为52.43%、84.47%和96.12%(P<0.01)。此外,电镜结果显示,400μg/mL的OPP对HUVECs作用48h能诱导细胞明显凋亡。结论OPP可通过抑制内皮细胞增殖、迁移、管腔形成和促进内皮细胞凋亡,而具有显著抗血管生成作用。

Background and objective Drugs which block tumor angiogenesis will be likely effective towards inhibiting tumor growth for angiogenesis being a prerequisite for tumor growth and metastasis. Therefore, antiangiogenesis has become a promising strategy for the treatment of cancer. Investigation on both antiangiogenic effect and mechanism（s） of oyster polypeptide （OPP） were performed via experiments of chicken embryos model in vivo and human umbilical vein endothelial cells （HUVECs） in vitro. Methods The methods employed in experiment were chorioallantoic membrane （CAM） angiogenesis in chicken embryos in vivo, MTT cell survival assay, flat plate scarification, transwell plates assay, matrigel-induced tube formation assay and transmission electron microscope et al. and the OPP＇s effects on angiogenesis was observed. Results Study showed that treatment with OPP resulted in significant inhibition of chorioallantoic membrane （CAM） angiogenesis in chicken embryos. MTT cell survival assay showed that treatment with OPP resulted in strong inhibition of HUVECs growth, with an IC50 of 400 μg/mL. Flat plate scarification suggested that OPP （200μg/mL, 400 μg/mL and 800μg/mL） distinctly inhibited HUVECs＇ migration （18.75%, 37.93%, 74.07% respectively, treatment for 12 h）. Treatment with OPP of different concentrations （200 μg/mL, 400 μg/mL and 800 μg/mL） significantly reduced the density of the migration cells by 15.5%, 37.2% and 67.24% （P〈0.05） respectively. Matrigel-induced tube formation assay showed that OPP resulted in striking inhibition of tube formation of 52.43%, 84.47% and 96.12% （P〈0.01） at 200μg/mL, 400μg/mL and 800μg/mL （treatment for 10 h） respectively. In addition, the apoptotic analysis by transmission electron microscope showed that OPP （400 μg/mL, treatment for 48 h） distinctly induced HUVECs＇ apoptosis. Conclusion This study strikingly showed that OPP could inhibit angiogenesis through its effects on vascular endothelial cells directly and the inhibition of their proliferation, migration, angiogenic ability, and induction of their apoptosis might be the antiangiogenic mechanism of OPP.