Pyruvate dehydrogenase kinases (PDKs) dominate the critical switch between mitochondria-based respiration and cytoplasm-based glycolysis by controlling pyruvate dehydrogenase (PDH) activity. Up-regulated PDKs play a great role in the Warburg effect in cancer cells and accordingly present a therapeutic target. Dichloroacetate (DCA) and AZD7545 are the two most-well-known PDK inhibitors exhibiting distinct pharmacological profiles. DCA showed anticancer effects in various preclinical models and clinical studies, while the primary preclinical indication of AZD7545 was on the improvement of glucose control in type II diabetes. Little, if any, study has been undertaken the elucidation of the effects of PDK inhibition on the metabolites in the tricarboxylic acid (TCA) cycle. Herein, the metabolite alterations of lung cancer cells (A549) upon the treatment with PDK inhibitors were studied using a reliable liquid-chromatography-based tandem mass spectrometry method. The developed method was validated for quantification of all common glycolysis and TCA cycle catabolites with good sensitivity and reproducibility, including glucose, pyruvate, lactate, acetyl coenzyme A, citrate, α-ketoglutarate, fumarate, succinate, malate, and oxaloacetate. Our results suggested that A549 cells exhibited distinct metabolite profiles following the treatment with DCA or AZD7545, which may reflect the different pharmacological indications of these two drugs.

中文翻译：

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液相色谱-串联质谱法揭示肺癌细胞在用不同的丙酮酸脱氢酶激酶抑制剂治疗后表现出不同的代谢产物特征

丙酮酸脱氢酶激酶（PDK）通过控制丙酮酸脱氢酶（PDH）的活性，主导了基于线粒体的呼吸作用与基于细胞质的糖酵解之间的关键转换。上调的PDK在癌细胞的Warburg效应中起重要作用，因此提出了治疗靶标。二氯乙酸盐（DCA）和AZD7545是两种最著名的PDK抑制剂，具有不同的药理作用。DCA在各种临床前模型和临床研究中均显示出抗癌作用，而AZD7545的主要临床前指征是改善II型糖尿病患者的血糖控制。很少有研究来阐明PDK抑制作用对三羧酸（TCA）循环中代谢产物的影响。在此处，使用可靠的基于液相色谱的串联质谱方法研究了用PDK抑制剂治疗后肺癌细胞（A549）的代谢物变化。验证了该开发方法可定量分析所有常见的糖酵解和TCA循环分解代谢物，具有良好的灵敏度和重现性，包括葡萄糖，丙酮酸，乳酸，乙酰辅酶A，柠檬酸，α-酮戊二酸，富马酸，琥珀酸，苹果酸和草酰乙酸。我们的结果表明，在用DCA或AZD7545处理后，A549细胞表现出独特的代谢产物特征，这可能反映了这两种药物的不同药理学适应症。验证了该开发方法可定量分析所有常见的糖酵解和TCA循环分解代谢物，具有良好的灵敏度和重现性，包括葡萄糖，丙酮酸，乳酸，乙酰辅酶A，柠檬酸，α-酮戊二酸，富马酸，琥珀酸，苹果酸和草酰乙酸。我们的结果表明，在用DCA或AZD7545处理后，A549细胞表现出独特的代谢产物特征，这可能反映了这两种药物的不同药理学适应症。验证了所开发的方法可定量分析所有常见的糖酵解和TCA循环分解代谢物，具有良好的灵敏度和重现性，包括葡萄糖，丙酮酸，乳酸，乙酰辅酶A，柠檬酸，α-酮戊二酸，富马酸，琥珀酸，苹果酸和草酰乙酸。我们的结果表明，在用DCA或AZD7545处理后，A549细胞表现出独特的代谢产物特征，这可能反映了这两种药物的不同药理学适应症。