Dichloroacetate and vitamin C synergistically suppress proliferation, migration and invasion of glioma U87 and U251 cells / 中国药理学通报

ABSTRACT

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 G1 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.