胡桃醌对胶质瘤细胞株U-87MG迁移、侵袭能力的抑制作用

Inhibitory effect of juglone on migration and invasion of glioma cell line U-87 MG

目的探讨肽基脯酰胺顺反异构酶Pin1(peptidyl-prolyl isomerase 1)抑制剂胡桃醌(Juglone)对胶质瘤细胞U-87 MG迁移、侵袭能力的影响并探讨相关机制。方法胶质瘤细胞株U-87 MG经0、0.8、1.6和3.2μmol·L-1胡桃醌处理后,以细胞划痕实验检测细胞迁移能力,transwell小室实验检测细胞侵袭能力,western blot法检测β-链蛋白(β-catenin)、血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)、基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)和基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)的蛋白表达水平。结果细胞划痕实验结果显示1.6μmol·L-1和3.2μmol·L-1胡桃醌组U-87 MG细胞划痕创面愈合率分别为46.04%±6.25%和30.05%±13.35%,与0μmol·L-1组比较明显下降(P<0.05);transwell小室实验结果显示1.6μmol·L-1和3.2μmol·L-1胡桃醌组穿膜细胞数平均为(103.67±5.69)个和(77.33±7.77)个,与0μmol·L-1组比较明显减少(P<0.05);western blot实验结果显示胡桃醌可显著下调U-87 MG细胞β-catenin、VEGF、MMP-2和MMP-9蛋白表达水平,作用随浓度增加而增大。结论胡桃醌可通过下调β-catenin及其下游MMP-2、MMP-9和VEGF的蛋白表达而抑制胶质瘤细胞迁移、侵袭能力。

ObjectiveTo investigate the effects and mechanisms of pin1（peptidyl-prolyl isomerase 1） inhibitor juglone on migration and invasion in glioma cell line U-87 MG.MethodsGlioma cells were treated with juglone at 0, 0.8,1.6 and 3.2 μmol·L-1. Wound-healing assay and invasion assay were performed to examine the inhibitory activity of juglone on glioma cell line U-87 MG. Western blot was used to analyze the protein expression of β-catenin, VEGF,MMP-2 and MMP-9.ResultsThe wound-healing assay showed that the wound-healing rate in juglone-treated groups was 46.04%±6.25% and 30.05%±13.35% at concentrations of 1.6 μmol·L-1and 3.2 μmol·L-1, repectively. Juglone treatment significantly reduced the wound-healing rate compared with controls（P 〈0.05）. Transwell invasion assay showed that the number of invaded cells in juglone–treated groups was 103.67±5.69 and 77.33±7.77 at the concentrations of1.6 μmol·L-1and 3.2 μmol·L-1, respectively. Juglone treatment significantly reduced cell invasion compared with controls（P 〈0.05）. Treatment with juglone significantly down-regulated the expression levels of β-catenin, VEGF, MMP-2and MMP-9 in U-87 MG cells in a dose-dependent manner.ConclusionThe present data suggests that juglone has a significant inhibitory action on cell migration and invasion through down-regulation of the β-catenin and its downstream VEGF, MMP-2 and MMP-9 protein expressions in glioma cell line U-87 MG.