二氯乙酸盐协同维生素C抑制神经胶质瘤细胞的增殖、迁移和侵袭及其作用机制  

Dichloroacetate and vitamin C synergistically suppress proliferation,migration and invasion of glioma U87 and U251 cells

作　　者：张晓宇 龙妮娅 刘健 出良钊 ZHANG Xiao-yu;LONG Ni-ya;LIU Jian;CHU Liang-zhao(School of Clinical Medicine,Guizhou Medical University;Dept of Neurosurgery Surgery of Guizhou Medical University;Guizhou Provincial People′s Hospital of Guizhou Medical University,Guiyang 550004,China)

机构地区：[1]贵州医科大学临床医学院 [2]贵州医科大学附属医院神经外科 [3]贵州省人民医院神经外科,贵州贵阳550004

出　　处：《中国药理学通报》2023年第2期275-286,共12页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金地区科学基金资助项目(No 81560409);贵州省神经系统疾病临床医学研究中心(No黔科合【2016】支撑2905)。

摘　　要：目的探讨二氯乙酸盐(dichloroacetate,DCA)与维生素C(vitamin C,VC)联合对胶质瘤U87和U251细胞恶性行为影响及机制。方法以不同浓度的DCA单用或联合5 mmol·L^(-1)的VC处理U87和U251细胞,CCK-8法检测各组增殖率后计算协同指数。以DMSO、15 mmol·L^(-1) DCA、5 mmol·L^(-1) VC及两者联合处理U87和U251细胞,体外实验检测各组克隆形成、活性氧含量、凋亡、周期、迁移和侵袭变化;皮下成瘤模型检测各组U251细胞体内增殖情况。应用网络药理分析DCA和VC共同作用靶点及其在胶质瘤中的价值后,Western blot检测各组细胞中BCL2A1和CDC25A的表达水平和降解率,以及CDK4、CDK6、cytochrome C、caspase-7和cleaved-caspase-7的表达水平。结果15 mmol·L^(-1) DCA与5 mmol·L^(-1) VC联合指数最大。与对照和单药组相比,联合组细胞体外克隆形成、迁移和侵袭能力明显降低,体内增殖率降低,活性氧含量、凋亡率和G 1期阻滞率明显增加。BCL2A1和CDC25A蛋白为DCA和VC在神经胶质瘤中的重要靶点;与对照和单药组相比,联合组细胞中的BCL2A1、CDC25A、CDK4和CDK6的表达量明显减少,Cytochrome C和cleaved-caspase-7的表达量明显增加,且联合组细胞中BCL2A1和CDC25A蛋白降解率明显增加。结论VC能够协同DCA促进BCL2A1和CDC25A的降解,抑制神经胶质瘤细胞的恶性行为。Aim To investigate the effects of dichloroacetate(DCA)combined with vitamin C(VC)on the malignant behavior of glioma U87 and U251 cells,and to explore the potential mechanism.Methods U87 and U251 cells were treated with different concentrations of DCA alone or in combination with 5 mmol·L^(-1) VC.The proliferation rate of each group was detected by CCK-8 method and the cooperative index was calculated.U87 and U251 cells were treated with DMSO,15 mmol·L^(-1) DCA,5 mmol·L^(-1) VC and their combination.The changes of clonal formation,reactive oxygen species content,apoptosis,cell cycle,migration and invasion were detected via in vitro experiments,while the proliferation of U251 cells in vivo in each group was detected by subcutaneous tumor-forming model.Western blot was used to detect the expression levels and degradation rates of BCL2A1 and CDC25A in each group of cells after network pharmacological analysis of DCA and VC targets and their value in glioma,and the expression levels of CDK4,CDK6,cytochrome C,caspase-7 and cleaved-caspase-7 were detected.Results The combined index of 15 mmol·L^(-1) DCA and 5 mmol·L^(-1) VC was the highest.Compared with the control and single drug groups,the clonal formation,migration and invasion ability of cells in combination group in vitro significantly decreased,the proliferation rate in vivo also decreased,and the content of reactive oxygen species,apoptosis rate and G 1 phase arrest rate significantly increased.BCL2A1 and CDC25A proteins were important targets of DCA and VC in glioma.Compared with the control and single-drug groups,the expression levels of BCL2A1,CDC25A,CDK4,and CDK6 in the combination group were significantly reduced,and the expression levels of cytochrome C and cleaved-caspase-7 markedly increased,and the protein degradation rates of BCL2A1 and CDC25A significantly increased in the combination group.Conclusions VC can cooperate with DCA to promote the degradation of BCL2A1 and CDC25A,and inhibit the malignant behavior of glioma cells.

关 键 词：二氯乙酸盐 维生素C 神经胶质瘤 协同 BCL2A1 CDC25A 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学] R730.264[医药卫生—基础医学] R739.41R977.23R977.6