常春藤皂苷元对U87 MG细胞生长增殖、迁移和侵袭的影响

Effect of hederagenin on growth, proliferation, migration and invasion of U87 MG cells

  • 摘要: 目的:探究常春藤皂苷元(Hed)对胶质母细胞瘤细胞系U87 MG 生长增殖以及运动能力的影响。方法:将Hed 分为0 μmol/L组、20 μmol/L组、40 μmol/L组、60 μmol/L组以及80 μmol/L组,采用CCK-8法检测Hed处理48 h的细胞活力以确定给药浓度;平板克隆实验检测细胞生长状况;EdU染色实验检测细胞DNA复制情况;流式细胞术检测细胞周期分布;伤口愈合实验和Transwell 实验检测细胞迁移和侵袭;蛋白免疫印迹实验检测细胞中Cyclin D1、CDK2、CDK4 以及MMP2 蛋白的表达。结果:Hed处理U87 MG细胞48 h后,40 μmol/L组的U87 MG细胞活力被显著抑制(P< 0.05)。与0 μmol/L组相比,20 μmol/L组和40 μmol/L组的U87 MG细胞的克隆形成率显著下降,DNA复制被显著抑制,G1期细胞所占比例显著减少,伤口愈合率显著下降,迁移和侵袭能力显著减弱(均P< 0.05),且抑制作用呈剂量依赖性。Cyclin D1、CDK2、CDK4以及MMP2蛋白的相对表达量均显著减少(均P< 0.05)。结论:Hed可抑制U87 MG胶质母细胞瘤的生长增殖和运动能力,其机制可能与抑制细胞周期相关蛋白以及MMP2有关。

     

    Abstract: Objective:To explore the effect of hederagenin(Hed)on the growth, proliferation and motor capacity of glioblastoma cell line U87 MG.Methods:Hed was divided into 0 μmol/L group, 20 μmol/L group, 40 μmol/L group, 60 μmol/L group and 80 μmol/L group.The cell viability of Hed treated for 48 h was detected by CCK-8 method to determine the drug administration concentration.Cell growth was detected by plate cloning assay.DNA replication was detected by EdU staining.Cell cycle distribution was detected by flow cytometry.Cell migration and invasion were detected by wound healing assay and Transwell assay.The expressions of Cyclin D1, CDK2, CDK4 and MMP2 proteins in cells were detected by western blotting.Results:After U87 MG cells were treated with Hed for 48 h, U87 MG cells in 40 μmol/L group were significantly inhibited (P< 0.05).Compared with 0 μmol/L group, the clonal formation rate of U87 MG cells in 20 μmol/L group and 40 μmol/L group significantly decreased, the DNA replication was significantly inhibited, the proportion of G1 phase cells was significantly reduced, the wound healing rate significantly decreased, and the migration and invasion ability were significantly weakened (P< 0.05), and the inhibition was dose-dependent.The relative expressions of Cyclin D1, CDK2, CDK4 and MMP2 proteins were significantlyreduced (P< 0.05).Conclusion:Hed can inhibit the growth, proliferation and motor capacity of U87 MG glioblastoma, and its mechanism may be related to the inhibition of cell cycle related proteins and MMP2.

     

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