脑型肌酸激酶促进胶质瘤干细胞增殖

Brain-type creatine kinase promotes proliferation of glioma stem cells

目的探讨脑型肌酸激酶(brain-type creatine kinase,CKB)对胶质瘤干细胞(glioma stem cells,GSCs)代谢和增殖能力的影响。方法应用代谢组学数据分析CKB的代谢底物肌酸在胶质瘤静脉与足背静脉血液样本中的含量差异;结合胶质母细胞瘤单细胞转录组测序数据分析、Western blot、qRT-PCR和免疫荧光染色等技术检测CKB在GSCs与非干性肿瘤细胞(non-stem tumor cells,NSTCs)中的表达差异;通过CCK-8实验检测敲低CKB或抑制剂环肌酸处理后GSCs和NSTCs的细胞增殖能力及磷酸肌酸对CKB敲低GSCs的细胞活力挽救能力。结果肌酸在胶质瘤静脉较足背静脉血液样本中的含量上升;在GSCs中CKB表达显著高于NSTCs;CKB敲低或环肌酸处理显著抑制GSCs的增殖能力,而对NSTCs影响较小;磷酸肌酸能在一定程度上挽救CKB敲低导致的GSCs活力下降。结论CKB在GSCs中表达上调,通过肌酸激酶/磷酸肌酸系统促进GSCs的增殖。

Objective To investigate the impact of brain-type creatine kinase(CKB)on the metabolism and proliferation capabilities of glioma stem cells(GSCs).Methods Metabolomics data was employed to analyze the difference in creatine levels,a metabolic substrate of CKB,between blood samples from glioma veins and pedal veins.Techniques such as single-cell transcriptome sequencing,Western blot,qRT-PCR,and immunofluorescence staining were used to detect the expression differences of CKB in GSCs compared to non-stem tumor cells(NSTCs).The CCK-8 assay was utilized to evaluate the proliferative capacity of GSCs and NSTCs after knocking down CKB or treating with a creatine cycle inhibitor.Additionally,the study assessed the ability of phosphocreatine to rescue the cell viability of GSCs with CKB knockdown.Results The concentration of creatine was found to be higher in blood samples from glioma veins than in pedal veins.CKB expression was significantly elevated in GSCs compared to NSTCs.Knocking down CKB or treating with a creatine cycle inhibitor significantly inhibited the proliferation of GSCs,with a lesser impact on NSTCs.Phosphocreatine was able to partially rescue the decrease in cell viability caused by CKB knockdown in GSCs.Conclusion CKB is upregulated in GSCs and enhances their proliferation through the creatine kinase/phosphocreatine system.