Department of Toxicology, School of Public Health, Peking University Health Science Center, Beijing 100191, PR China; Beijing Key Laboratory of Toxicological Research and Risk Assessment for Food Safety, School of Public Health, Peking University Health Science Center, Beijing 100191, PR China.
National Institute of Environmental Health, Chinese Center for Disease Control and Prevention, Beijing 100021, PR China.
Toxicol Appl Pharmacol. 2021 Sep 1;426:115640. doi: 10.1016/j.taap.2021.115640. Epub 2021 Jul 7.
Benzo[a]pyrene (BaP) is a strong carcinogen for lung cancer, and forkhead-box A1 (FOXA1) plays an oncogenic role in BaP-transformed cell THBEc1. To explore the remodeling of metabolic pattern caused by BaP-induced transformation and the possible role FOXA1 might play in it, we compared metabolic patterns between THBEc1 cells and control using untargeted metabolomics and lipidomics analysis, and determined the effects of FOXA1 knockout on the metabolic pattern of THBEc1 cells. Metabolomics and lipidomics identified a total of 15 and 46 differential metabolites and lipids between THBEc1 and 16HBE cells, respectively, and a total of 4 and 1 differential metabolites and lipids between FOXA1 knockout cell THBEc1-ΔFOXA1-c34 and control cell THBEc1-ctrl, respectively. Analysis results of metabolites and metabolic pathways indicated the metabolic pattern remodeling may be related to the alteration in glucose metabolism during BaP-induced transformation. Western blotting revealed the up-regulation of enolase-2 (ENO2), pyruvate carboxylase (PCB), aconitase-2 (ACO2) and phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2) (Thr202/Tyr204), the down-regulation of succinate dehydrogenase complex subunit A (SDHA) and phosphoenolpyruvate carboxykinase 2 (PCK2) in THBEc1 cells. The detection results of metabolites related to glucose metabolism demonstrated the decreasing of lactic acid content in cells, lactic acid production in culture medium and citric acid content in mitochondria, and the increasing of ATP production of THBEc1 cells. FOXA1 knockout partially reversed the changes of ENO2, SDHA, PCK2 and p-ERK1/2 (Thr202/Tyr204) levels, lactic acid release, citric acid content in mitochondria of THBEc1 cells. In conclusion, FOXA1 knockout partially reversed the remodeling of glucose metabolism caused by BaP-induced malignant transformation. Our findings provide a clue for the possible role of FOXA1 in glucose metabolism regulation.
苯并[a]芘（BaP）是肺癌的强致癌物，叉头框转录因子 A1（FOXA1）在 BaP 转化的 THBEc1 细胞中发挥致癌作用。为了探索 BaP 诱导转化引起的代谢模式重塑以及 FOXA1 可能在其中发挥的作用，我们使用非靶向代谢组学和脂质组学分析比较了 THBEc1 细胞和对照细胞之间的代谢模式，并确定了 FOXA1 敲除对 THBEc1 细胞代谢模式的影响。代谢组学和脂质组学分别在 THBEc1 和 16HBE 细胞之间鉴定出 15 种和 46 种差异代谢物和脂质，在 FOXA1 敲除细胞 THBEc1-ΔFOXA1-c34 和对照细胞 THBEc1-ctrl 之间分别鉴定出 4 种和 1 种差异代谢物和脂质。代谢物和代谢途径的分析结果表明，代谢模式重塑可能与 BaP 诱导转化过程中葡萄糖代谢的改变有关。Western blot 显示，THBEc1 细胞中烯醇化酶 2（ENO2）、丙酮酸羧化酶（PCB）、 aconitase-2（ACO2）和磷酸化细胞外信号调节激酶 1/2（p-ERK1/2）（Thr202/Tyr204）的上调，琥珀酸脱氢酶复合体亚单位 A（SDHA）和磷酸烯醇丙酮酸羧激酶 2（PCK2）的下调。葡萄糖代谢相关代谢物的检测结果表明，THBEc1 细胞内乳酸含量降低，培养基中乳酸生成减少，线粒体中柠檬酸含量增加，THBEc1 细胞 ATP 生成增加。FOXA1 敲除部分逆转了 THBEc1 细胞中 ENO2、SDHA、PCK2 和 p-ERK1/2（Thr202/Tyr204）水平、乳酸释放、线粒体中柠檬酸含量的变化。总之，FOXA1 敲除部分逆转了 BaP 诱导的恶性转化引起的葡萄糖代谢重塑。我们的研究结果为 FOXA1 在葡萄糖代谢调节中的可能作用提供了线索。
