蒲葵子乙醇提取物对人肝癌Bel-7402细胞增殖和迁移的影响

Effects of Livistona chinensis seeds on proliferation and migration of hepatocellular carcinoma Bel-7402 cells

目的研究蒲葵子乙醇提取物(EELC)对人肝癌Bel-7402细胞增殖和迁移的影响。方法将培养的Bel-7402细胞随机分为5组:对照组、150μg/ml EELC处理组、300μg/ml EELC处理组、10μmol/L Notch抑制剂DAPT处理组和10μg/ml抗血管内皮生长因子(VEGF)抗体处理组,分别处理后,采用MTT和划痕实验检测各组细胞增殖和迁移能力的变化,Western blot检测各组细胞内VEGF、基质金属蛋白酶-2(MMP-2)及Notch胞内片段(NICD)等蛋白表达的变化,ELISA检测各组细胞分泌VEGF水平的变化。结果不同浓度EELC、DAPT和抗VEGF抗体处理Bel-7402细胞后,Notch-VEGF途径相关的VEGF、MMP-2、NICD等蛋白表达水平、VEGF分泌量、细胞的增殖和迁移能力均受到不同程度地抑制(P<0.05)。而且EELC的抑制作用呈现一定的浓度依赖性和时间依赖性。结论蒲葵子乙醇提取物可通过Notch-VEGF途径抑制人肝癌Bel-7402细胞的增殖和迁移。

【Objective】 To study the effect of ethanol extract of Livistona chinensis seeds（EELC） on the proliferation and migration of human hepatocellular carcinoma Bel-7402 cells. 【Methods】 Cultured Bel-7402 cells were randomized into five groups： control group, 150 μg/ml EELC group, 300 μg/ml EELC group, 10μmol/L DAPT group and 10 μg/ml anti-vascular endothelial growth factor（VEGF） group. After treatment, cell proliferation was determined by MTT assay and cell migration by wound-healing assay. Furthermore, the expressions of VEGF, matrix metalloproteinase-2（MMP-2） and Notch intracellular domain（NICD） in Bel-7402 cells were evaluated by Western blot, and the secretion of VEGF protein was detected by ELISA. 【Results】Treatments with different concentrations of DAPT, anti-VEGF antibody and EELC inhibited the proliferation and migration of Bel-7402 cells, as well as the expressions of VEGF, MMP-2 and NICD proteins（P〈0.05）.Additionally, EELC inhibited cell proliferation and migration as well as gene expressions in a dose- and time-dependent manner. 【Conclusion】 EELC may inhibit the proliferation and migration of human hepatocellular carcinoma Bel-7402 cells through Notch-VEGF pathway.