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目的探究隐丹参酮(CTS)对U2OS骨肉瘤细胞株血管内皮生长因子(VEGF)的调控作用以及对血管生成的影响。方法体外培养的U2OS骨肉瘤细胞分为CTS处理组和对照组,处理组给予(20、40、80、160)μmol/L CTS,对照组给予等体积培养液。实时荧光定量PCR检测U2OS细胞VEGF mRNA的变化,Western blot法和免疫荧光细胞化学染色技术检测VEGF蛋白表达,CCK-8法检测CTS对细胞生长的影响,体外成管实验观察血管形成情况。结果 CTS可明显抑制U2OS细胞内VEGF mRNA和蛋白的表达,且该抑制效应具有剂量依赖性。CCK-8法实验结果显示CTS能抑制U2OS细胞的生长。体外成管实验结果证明CTS对新生血管的形成有抑制作用。结论 CTS可下调U2OS骨肉瘤细胞VEGF的水平,并能抑制血管的生成。
Objective To investigate the regulatory effect of cryptotanshinone (CTS) on vascular endothelial growth factor (VEGF) in U2OS osteosarcoma cell line and its effect on angiogenesis. Methods U2OS osteosarcoma cells cultured in vitro were divided into CTS treatment group and control group. The treatment group was given CTS (20, 40, 80, 160) μmol / L, and the control group was given equal volume of culture medium. The changes of VEGF mRNA in U2OS cells were detected by real-time fluorescence quantitative PCR. The expression of VEGF protein was detected by Western blot and immunofluorescence cytochemistry. The effect of CTS on cell growth was detected by CCK-8 assay. Results CTS significantly inhibited the expression of VEGF mRNA and protein in U2OS cells, and the inhibitory effect was dose-dependent. CCK-8 test results show that CTS can inhibit the growth of U2OS cells. In vitro tube formation test results show that CTS has an inhibitory effect on the formation of new blood vessels. Conclusion CTS can down-regulate the VEGF level in U2OS osteosarcoma cells and inhibit angiogenesis.