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目的探讨和厚朴酚(HNK)对人白血病U937细胞侵袭及血管新生作用的影响及其可能的分子机制。方法应用不同浓度的HNK处理U937细胞不同时间,用MTT法检测HNK对细胞增殖抑制作用;基质黏附试验检测HNK对U937细胞黏附功能的影响;TranswellTM小室检测HNK对U937细胞侵袭抑制作用。实时定量PCR(qRT-PCR)检测血管内皮生长因子(VEGF)、血管内皮生长因子受体1(VEGFR1)、基质金属蛋白酶9(MMP-9)mRNA水平变化。ELISA检测HNK处理U937细胞后上清VEGF蛋白表达水平。结果 HNK对U937的体外增殖有显著抑制作用,呈剂量和时间依赖性。低浓度HNK能够抑制U937细胞的黏附及侵袭能力。不同浓度HNK处理U937细胞24 h后,细胞VEGF、VEGFR1及MMP-9表达水平呈剂量依赖性下调。结论 HNK可能通过抑制VEGF、VEGFR1及MMP-9表达,抑制U937细胞侵袭及血管新生功能。
Objective To investigate the effect of honokiol (HNK) on the invasion and angiogenesis of human leukemia U937 cells and its possible molecular mechanism. Methods U937 cells were treated with different concentrations of HNK for different time. MTT assay was used to detect the effect of HNK on the proliferation of U937 cells. Matrix adhesion assay was used to detect the effect of HNK on the adhesion of U937 cells. TranswellTM was used to detect the inhibitory effect of HNK on U937 cells. Real-time quantitative PCR (qRT-PCR) was used to detect the changes of vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 1 (VEGFR1) and matrix metalloproteinase 9 (MMP-9) The expression of VEGF protein in supernatant of U937 cells treated with HNK was detected by ELISA. Results HNK significantly inhibited the proliferation of U937 cells in a dose-and time-dependent manner. Low concentration of HNK can inhibit the adhesion and invasion of U937 cells. The expression of VEGF, VEGFR1 and MMP-9 in U937 cells treated with different concentrations of HNK for 24 h were down-regulated in a dose-dependent manner. Conclusion HNK may inhibit the invasion and angiogenesis of U937 cells by inhibiting the expression of VEGF, VEGFR1 and MMP-9.