华蟾素注射液干预血管内皮细胞和肿瘤细胞增殖及VEGF表达的实验研究

摘要

目的 观察华蟾素对人脐静脉内皮细胞 (HUVEC) 和人结肠癌SW480细胞的抑制作用和VEGF表达的影响, 探讨华蟾素抑制肿瘤血管生成的可能作用机制.方法 体外分别培养两种细胞, MTT法检测不同浓度华蟾素在不同作用时间对两种细胞的生长抑制, 倒置荧光显微镜实时成像技术观察细胞凋亡形态, Elisa法检测华蟾素对两种细胞表达VEGF及VEGFR-2含量的影响.结果华蟾素能够有效抑制肿瘤细胞生长, 不能诱导正常血管内皮细胞的凋亡;华蟾素有效降低SW480细胞VEGF含量和HUVEC细胞膜VEGFR-2受体的表达, 但对HUVEC细胞VEGF无明显抑制作用.结论 华蟾素能够选择性抑制肿瘤细胞生长和增殖, 同时不会引起正常血管细胞的损伤, 华蟾素抑制肿瘤新生血管生成的作用机理可能是阻断了肿瘤细胞所表达VEGF与正常内皮细胞膜表面的VEGFR-2受体结合.%ObjectiveTo explore the mechanism of Cinobufacini inhibiting the tumor angiogenesis by observing the effect of Cinobufacini injection on the growth inhibition of human umbilical vein endothelial cell (HUVEC) and human colon carcinoma SW480 as well as the expression of VEGF protein.MethodsHUVEC and SW480 cells were cultured in vitro with MTT method for detection of two kinds of cells growth inhibitions in different concentrations and Real-time imaging technique for the morphology of cellular apoptosis.Elisa was used to detect the expression of VEGF and VEGF receptors.Results CINO refrained effectively the growth of SW480 cells, but it failed to induce the apoptosis of HUVEC cells;CINO decreased VEGF and VEGFR-2 levels in SW480 cells, VEGFR-2 level in HUVEC (P＜0.01), but it showed no obvious refrain effect on VEGF level in HUVEC cells.Conclusion Our results show that CINO has anti-cancer activities while protects normal blood vessels from being injured, and the possible mechanism of how CINO inhibits tumor neovascularization-CINO interdicts the combination between VEGF expressed by tumor cells and its receptor VEGFR-2 embedded in the membranes of normal endothelial cells.