小分子化合物小檗碱靶向CDK2抑制C6神经胶质瘤细胞生长的分子机制研究

Molecular Mechanism for Small Molecule Berbine Inhibiting C6 Proliferation by Targeting Cycline-dependent Kinase-2

目的探究天然小分子化合物小檗碱对神经胶质瘤的抑制作用。方法选择神经胶质瘤C6细胞株为实验模型,通过噻唑蓝(MTT)比色法实验检测0、1、5、10、25、50μmol/L的小檗碱对C6细胞增殖的抑制作用,采用细胞划痕实验验证其抑制C6细胞的迁移能力,利用平板克隆实验验证小檗碱抑制C6细胞的克隆形成能力,并通过免疫蛋白印迹法检测不同浓度的小檗碱对细胞的CDK2蛋白表达水平的影响。以CDK2为靶点,进行小檗碱与CDK2分子对接。结果小檗碱明显抑制C6细胞的增殖、迁移与克隆形成。并且,小檗碱能够下调CDK2的蛋白表达。小檗碱能够与CDK2的ATP结合区结合。结论小檗碱可能靶向CDK2,下调CDK2的表达从而抑制C6细胞的生长。

Objective To explore the inhibition of neuroglioma by using a natural small molecular compound berberine.Methods C6 glioma cell line was selected as the model.The inhibition of C6 proliferation via different concentration of berbine 0,1,5,10,25,50μmol/L were detected by MTT assay and cell colony formation.The suppression effect on the migration was verified by cell wound test.And the expression of CDK2 was detected by western blot.Computer simulation was performed based on the CDK2 and berbine.Results Berberine could significantly inhibit the proliferation,migration and colony formation of C6 cells.And CDK2 was downregulated after treatment with berbine.Berbine could bind to the ATP region of CDK2.Conclusion Berbine could inhibit the proliferation of C6 glioma by targeting CDK2 and downregulating CDK2 as well.