Vo Vu T A, Choi Jong-Whan, Phan Ai N H, Hua Tuyen N M, Kim Min-Kyu, Kang Byoung Heon, Jung Soon-Hee, Yong Suk-Joong, Jeong Yangsik
Department of Biochemistry, Yonsei University Wonju College of Medicine, Wonju, Republic of Korea.
Department of Global Medical Science, Medical Research Center, Institute of Lifestyle Medicine, Yonsei University Wonju College of Medicine, Wonju, Republic of Korea.
Anticancer Res. 2018 Apr;38(4):2187-2193. doi: 10.21873/anticanres.12460.
BACKGROUND/AIM: Cancer cells are distinct in terms of glutamine dependence. Here we investigated the different susceptibility of glutamine-independent and glutamine-dependent non-small cell lung cancer (NSCLC) to treatment with tumor necrosis factor receptor-associated protein 1 (TRAP1) inhibitor gamitrinib-triphenylphosphonium (G-TPP).
Cell viability and proliferation under glutamine deprivation and G-TPP treatment were determined by the MTT and colony-formation assays. Protein and mRNA expression were determined by western blot and quantitative polymerase chain reaction. Colorimetric-based assay was performed to check for glutamine synthetase (GS) activity.
NSCLC cells showed diverse adaptation under glutamine-depleted condition and were categorized into glutamine-independent and glutamine-dependent cells. Treatment with G-TPP particularly increased GS activity and induced cell death due to energy shortage indicated by phosphorylated AMP-activated protein kinase (AMPK) in glutamine-dependent cells.
This finding provides better understanding of TRAP1-mediated glutamine metabolism through GS activity, and evidence that TRAP1 could be a promising therapeutic target for glutamine-addicted cancer.
背景/目的：癌细胞在谷氨酰胺依赖性方面存在差异。在此，我们研究了不依赖谷氨酰胺和依赖谷氨酰胺的非小细胞肺癌（NSCLC）对肿瘤坏死因子受体相关蛋白1（TRAP1）抑制剂加米替尼-三苯基膦（G-TPP）治疗的不同敏感性。
通过MTT和集落形成试验测定谷氨酰胺剥夺和G-TPP处理下的细胞活力和增殖。通过蛋白质印迹和定量聚合酶链反应测定蛋白质和mRNA表达。进行比色法检测谷氨酰胺合成酶（GS）活性。
NSCLC细胞在谷氨酰胺缺乏条件下表现出不同的适应性，并被分为不依赖谷氨酰胺和依赖谷氨酰胺的细胞。在依赖谷氨酰胺的细胞中，G-TPP处理特别增加了GS活性，并因能量短缺诱导细胞死亡，这由磷酸化的AMP激活蛋白激酶（AMPK）表明。
这一发现有助于更好地理解TRAP1通过GS活性介导的谷氨酰胺代谢，并证明TRAP1可能是对谷氨酰胺成瘾癌症的一个有前景的治疗靶点。
