ABSTRACT

Elucidating the dysregulated metabolic pathways in cancer cells and their relevance to cisplatin resistance could yield new insights into cancer therapy. We previously reported that eight metabolites involved in the tricarboxylic acid (TCA) cycle and glutamine metabolism were associated with platinum-based chemotherapy efficacy in human lung cancer. Here, we investigated the metabolic differences upon cisplatin treatment in lung cancer in vitro and in vivo. A simple and partially validated standard addition method was applied for the quantification of five metabolites involved in the TCA cycle and glutamine metabolism using amide hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS). The present study investigated the levels of these biomarkers in A549 cells and the cisplatin-resistant A549-DDP cells, as well as in the plasma before and after cisplatin treatment in A549 xenograft mice. Levels of five metabolites, including 2-hydroxyglutaric acid (2-HG), α-ketoglutarate (α-KG), succinate, glutamine, and glutamate, showed a decreasing trend in A549-DDP cells. In addition, 2-HG and glutamine were the most significantly altered metabolites in cisplatin-treated A549 xenograft mice. These data indicate that the TCA cycle and glutamine metabolism play important roles in cisplatin-based chemotherapy resistance in lung cancer. Our results provide a new angle for exploring the molecular mechanisms underlying cisplatin resistance.

摘要

阐明癌细胞中失调的代谢途径及其与顺铂耐药的相关性可以为癌症治疗带来新的见解。我们之前报道了参与三羧酸 (TCA) 循环和谷氨酰胺代谢的八种代谢物与人肺癌中基于铂的化疗疗效相关。在这里，我们研究了顺铂在体外和体内治疗肺癌时的代谢差异。使用酰胺亲水相互作用液相色谱-串联质谱 (HILIC-MS/MS)，采用简单且部分验证的标准添加方法对参与 TCA 循环和谷氨酰胺代谢的五种代谢物进行定量。本研究调查了这些生物标志物在 A549 细胞和顺铂耐药的 A549-DDP 细胞中的水平，以及在 A549 异种移植小鼠顺铂治疗前后的血浆中。A549-DDP 细胞中 2-羟基戊二酸 (2-HG)、α-酮戊二酸 (α-KG)、琥珀酸、谷氨酰胺和谷氨酸等五种代谢物的水平呈下降趋势。此外，在顺铂治疗的 A549 异种移植小鼠中，2-HG 和谷氨酰胺是最显着改变的代谢物。这些数据表明，TCA 循环和谷氨酰胺代谢在肺癌中以顺铂为基础的化疗耐药中起重要作用。我们的结果为探索顺铂耐药的分子机制提供了一个新的角度。A549-DDP 细胞呈下降趋势。此外，在顺铂治疗的 A549 异种移植小鼠中，2-HG 和谷氨酰胺是最显着改变的代谢物。这些数据表明，TCA 循环和谷氨酰胺代谢在肺癌中以顺铂为基础的化疗耐药中起重要作用。我们的结果为探索顺铂耐药的分子机制提供了一个新的角度。A549-DDP 细胞呈下降趋势。此外，在顺铂治疗的 A549 异种移植小鼠中，2-HG 和谷氨酰胺是最显着改变的代谢物。这些数据表明，TCA 循环和谷氨酰胺代谢在肺癌中以顺铂为基础的化疗耐药中起重要作用。我们的结果为探索顺铂耐药的分子机制提供了一个新的角度。