EFFECT OF LACTATE DEHYDROGENASE INHIBITION BY OXAMATE ON LEWIS LUNG CARCINOMA CELLS WITH DIFFERENT METASTATIC POTENTIAL
Keywords:lactate dehydrogenase inhibitor, oxamate, cancer cells, antimetastatic therapy
Background. Today, the ability for metabolic reprogramming is considered one of the distinguishing features of metastatically active tumor cells, a classic example of which is aerobic glycolysis. Despite a large number of studies in this direction, the question of the relationship between the intensity of aerobic glycolysis and the metastatic potential of tumor cells remains almost completely open. The work aimed to investigate the effect of the lactate dehydrogenase (LDH) inhibitor on the viability and several characteristics of Lewis lung carcinoma cells with different metastatic potential. Materials and Methods. High-metastatic (LLC) and low-metastatic (LLC/R9) variants of Lewis lung carcinoma cells were used. After 24 h of tumor cells incubation with or without 40 mM sodium oxamate, cell viability, the concentration of glucose and lactate in the incubation medium, distribution of cells by the cell cycle phases, and intracellular ROS production were estimated. Results. It was revealed that regardless of the metastatic potential, LLC cells are heterogeneous in terms of both the involvement of aerobic glycolysis in their growth and survival processes and the sensitivity to the cytotoxic/cytostatic action of an LDH inhibitor. 35% of cells of either LLC variant form an oxamate-resistant subpopulation while 65% are oxamate-sensitive. The rate of glucose consumption of LLC/R9 cells in the absence of oxamate is almost twice higher compared to LLC and, as a result, the sensitivity of these cells to the cytotoxic/cytostatic effect of oxamate also is significantly higher (the IC50 for LLC/R9 cells is by 35.8% lower than that for LLC cells, p < 0.05). Approximately one-third of the cells of both LLC and LLC/R9 variants can survive and proliferate when aerobic glycolysis is completely inhibited by oxamate. This indicates metabolic reprogramming (either pre-existing or dynamically arising in response to inhibition of glycolysis) of this subpopulation of cells, within which not only the survival of cells but also their proliferative activity is most likely based on glutamine metabolism. Conclusions. Such metabolic heterogeneity of metastatically active cells indicates that inhibition of glycolysis as monotherapy is insufficient for effective antimetastatic therapy. Presumably, more effective would be to involve various inhibitors of metabolic processes that ensure the metabolic plasticity of metastatic cells.
Weber GF. Time and circumstances: cancer cell metabolism at various stages of disease progression. Front Oncol. 2016;6:257. doi: 10.3389/fonc.2016.00257.
Lloyd MC, Cunningham JJ, Bui MM, et al. Darwinian dynamics of intratumoral heterogeneity: Not solely ran- dom mutations but also variable environmental selection forces. Cancer Res. 2016;76:3136-3144.
Chen J, Cao S, Si-Tu B, et al. Metabolic reprogramming-based characterization of circulating tumor cells in prostate cancer. J Exp Clin Cancer Res. 2018;37:127.
Schild T, Low V, Blenis J, et al. Unique metabolic adaptations dictate distal organ-specific metastatic colonization.
Cancer Cell. 2018;33;347-354.
Ohshima K, Morii E. Metabolic reprogramming of cancer cells during tumor progression and metastasis. Me tabolites. 2021;11(1):28. doi: 10.3390/metabo11010028
Roda N, Gambino V, Giorgio M. Metabolic constrains rule metastasis progression. Cells. 2020;9:2081. doi:10.3390/ cells9092081
Warburg O. On the origin of cancer cells. Science. 1956;123:309-314.
Vaupel P, Schmidberger H, Mayer A. The Warburg effect: essential part of metabolic reprogramming and central contributor to cancer progression. Int J Radiat Biol. 2019;95:912-919. doi: 10.1080/09553002.2019.1589653
Boedtkjer E, Pedersen SF. The acidic tumor microenvironment as a driver of cancer. Annu Rev Physiol. 2020;82:103–126.
Gandhi N, Das GM. Metabolic reprogramming in breast cancer and its therapeutic implications. Cells. 2019;8:89. doi: 10.3390/cells8020089
Nagao A, Kobayashi M, Koyasu S, et al. HIF-1-dependent reprogramming of glucose metabolic pathway of cancer cells and its therapeutic significance. Int J Mol Sci. 2019;20:238. doi: 10.3390/ijms20020238
Ward PS, Thompson CB. Metabolic reprogramming: a cancer hallmark even Warburg did not anticipate. Cancer Cell. 2012;21:297-308.
Tasdogan A, Faubert B, Ramesh V, et al. Metabolic heterogeneity confers differences in melanoma metastatic potential. Nature. 2019;577:115-120. doi: 10.1038/s41586-019-1847-2
Vaupel P, Multhoff G. The Warburg effect: historical dogma versus current rationale. Adv Exp Med Biol. 2021;1269:169-177. doi: 10.1007/978-3-030-48238-1_27
Pyaskovskaya ON, Kolesnik DL, Garmanchouk LV, et al. Role of tumor/endothelial cell interactions in tu- mor growth and metastasis. Exp Oncol. 2021;43(2):104-110. doi: 10.32471/exp-oncology.2312-8852.vol-43- no-2.16157
Nicoletti I, Migliorati G, Pagliacci MC, et al. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods. 1991;139:271-280. doi: 10.1016/0022- 1759(91)90198-o
Wang H, Joseph JA. Quantifying cellular oxidative stress by dichlorofluorescein assay using microplate reader.
Free Radic Biol Med. 1999;27:612-616. doi: 10.1016/s0891-5849(99)00107-0
Peng Y, Fu S, Hu W, et al. Glutamine synthetase facilitates cancer cells to recover from irradiation-induced G2/M arrest. Cancer Biol Ther. 2020;21(1):43-51. doi: 10.1080/15384047.2019.1665394
Zhang W, Wang C, Hu X, et al. Inhibition of LDHA suppresses cell proliferation and increases mitochondrial apoptosis via the JNK signaling pathway in cervical cancer cells. Oncol Rep. 2022;47(4):77. doi: 10.3892/or.2022.8288
Lyssiotis C, Son J, Mancias J, et al. Pancreatic cancers depend on a non-canonical glutamine metabolism pathway.
Cancer Metab. 2014;2:44. doi: 10.1186/2049-3002-2-S1-P44
Scalise M, Pochini L, Galluccio M, et al. Glutamine transport and mitochondrial metabolism in cancer cell growth. Front Oncol. 2017;7:306. doi: 10.3389/fonc.2017.00306
Mori S, Chang J, Andrechek E, et al. Anchorage-independent cell growth signature identifies tumors with metastatic potential. Oncogene. 2009;28:2796-2805. doi:10.1038/onc.2009.139
LeBleu VS, O’Connell JT, Gonzalez Herrera KN, et al. PGC-1α mediates mitochondrial biogenesis and oxidative phosphorylation in cancer cells to promote metastasis. Nat Cell Biol. 2014;16(10):992-1003. doi: 10.1038/ncb3039
Tang E, Liu S, Zhang Z, et al. Therapeutic potential of glutamine pathway in lung cancer. Front Oncol. 2022;11:835141. doi: 10.3389/fonc.2021.835141
Halama A, Suhre K. Advancing cancer treatment by targeting glutamine metabolism-a roadmap. Cancers (Basel). 2022;14(3):553. doi: 10.3390/cancers14030553
Boudreau A, Purkey HE, Hitz A, et al. Metabolic plasticity underpins innate and acquired resistance to LDHA inhibition. Nat Chem Biol. 2016;12(10):779-786. doi: 10.1038/nchembio.2143
Submitted: November 11, 2022
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