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目的从美国肉参(Isostichopus badionotus)中分离纯化岩藻糖基化海参硫酸软骨素(sea cucumber chondroitin sulfate,SC-CHS),体外研究其对肿瘤诱导血管生成的抑制作用。方法采用MTT法检测SC-CHS对95D细胞增殖活性的影响;小管形成实验研究SC-CHS对脐静脉内皮细胞(HUVEC)小管形成能力的抑制作用;采用鸡胚尿囊膜(CAM)新生血管模型,研究其对血管生成的影响;通过RT-PCR和WesternBlot法检测其对肿瘤诱导血管生成相关因子缺氧诱导因子(hypoxia inducible factor-1α,Hif-1α)和血管内皮生长因子(vascular endothelial growth factor,VEGF)mRNA和蛋白的表达。结果结果显示SC-CHS能显著抑制95D细胞的增殖,能抑制HUVEC的小管形成作用和CAM血管新生。RT-PCR和Western Blot检测结果表明,高剂量SC-CHS能显著降低95D细胞中Hif-1α和VEGF mRNA的表达,减少VEGF蛋白的合成。结论SC-CHS体外能显著抑制肿瘤血管生成,这可能是通过影响肿瘤细胞中VEGF的合成发挥作用。
Objective To isolate and purify sea cucumber chondroitin sulfate (SC-CHS) from American ginseng (Isostichopus badionotus) and study its inhibitory effect on tumor-induced angiogenesis in vitro. Methods The effect of SC-CHS on the proliferation of 95D cells was detected by MTT assay. The inhibitory effect of SC-CHS on the tubular formation of human umbilical vein endothelial cells (HUVEC) The effects of hypoxia inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) on angiogenesis were analyzed by RT-PCR and Western Blot. , VEGF) mRNA and protein expression. The results showed that SC-CHS could significantly inhibit the proliferation of 95D cells, inhibit the tubular formation of HUVEC and CAM angiogenesis. The results of RT-PCR and Western Blot showed that high-dose SC-CHS could significantly decrease the expression of Hif-1α and VEGF mRNA and decrease the VEGF protein synthesis in 95D cells. Conclusion SC-CHS can significantly inhibit tumor angiogenesis in vitro, which may play a role in the synthesis of VEGF in tumor cells.