Inhibition of LDH-A by oxamate induces G2/M arrest, apoptosis and increases radiosensitivity in nasopharyngeal carcinoma cells

Zhai,Xiaoming; Yang,Yang; Wan,Jianmei; Zhu,Ran; Wu,Yiwei

doi:10.3892/or.2013.2735

Inhibition of LDH-A by oxamate induces G2/M arrest, apoptosis and increases radiosensitivity in nasopharyngeal carcinoma cells

Authors:
- Xiaoming Zhai
- Yang Yang
- Jianmei Wan
- Ran Zhu
- Yiwei Wu
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Affiliations: Department of Radiation Oncology, The First Affiliated Hospital, Soochow University, Suzhou, P.R. China, Department of Radiation Therapy, Zhejiang Cancer Hospital, Hangzhou, Zhejiang, P.R. China, School of Radiation Medicine and Protection, Medical College, Soochow University, Suzhou, Jiangsu 215006, P.R. China, Department of Nuclear Medicine, The First Affiliated Hospital, Soochow University, Suzhou, Jiangsu 215006, P.R. China
Published online on: September 19, 2013 https://doi.org/10.3892/or.2013.2735
Pages: 2983-2991

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Abstract

An elevated rate of glucose consumption and the dependency on aerobic glycolysis for ATP generation have long been observed in cancer cells, a phenomenon known as the Warburg effect. the altered energy metabolism in cancer cells provides an attractive opportunity for developing novel cancer therapeutic strategies. Lactate dehydrogenase (LDH), which catalyzes the transformation of pyruvate to lactate, plays a vital role in the process of glycolysis. It has been reported that the level of LDH-A expression is increased both in head and neck cancer cells and in the blood serum of nasopharyngeal carcinoma (NPC) patients, and is associated with poor prognosis. However, the effect of LDH-A inhibition on NPC cells remains unknown. Here, in the present study, we found that oxamate, a classical inhibitor of LDH-A, suppressed cell proliferation in a dose- and time-dependent manner both in CNE-1 and CNE-2 cells, two NPC cancer cell lines. LDH inhibition by oxamate induced G2/M cell cycle arrest via downregulation of the CDK1/cyclin B1 pathway and promoted apoptosis through enhancement of mitochondrial ROS generation. N-acetylcysteine, a specific scavenger of ROS, significantly blocked the growth inhibition effect induced by oxamate. We also identified that oxamate increased sensitivity to ionizing radiation in the two NPC cancer cell lines. Furthermore, we verified similar results in tumor xenograft models. collectively, these results suggest that LDH-A may serve as a promising therapeutic target for NPC treatment.

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1	Wei WI and Sham JS: Nasopharyngeal carcinoma. Lancet. 365:2041–2054. 2005. View Article : Google Scholar : PubMed/NCBI
2	Guigay J: Advances in nasopharyngeal carcinoma. Curr Opin Oncol. 20:264–269. 2008. View Article : Google Scholar : PubMed/NCBI
3	Xie P, Yue JB, Fu Z, Feng R and Yu JM: Prognostic value of ¹⁸F-FDG PET/CT before and after radiotherapy for locally advanced nasopharyngeal carcinoma. Ann Oncol. 21:1078–1082. 2010.
4	Chan SC, Chang JT, Wang HM, et al: Prediction for distant failure in patients with stage M0 nasopharyngeal carcinoma: the role of standardized uptake value. Oral Oncol. 45:52–58. 2009. View Article : Google Scholar : PubMed/NCBI
5	Pelicano H, Martin DS, Xu RH and Huang P: Glycolysis inhibition for anticancer treatment. Oncogene. 25:4633–4646. 2006. View Article : Google Scholar : PubMed/NCBI
6	Goldman RD, Kaplan NO and Hall TC: Lactic dehydrogenase in human neoplastic tissues. Cancer Res. 24:389–399. 1964.PubMed/NCBI
7	Fantin VR, St-Pierre J and Leder P: Attenuation of LDH-A expression uncovers a link between glycolysis, mitochondrial physiology, and tumor maintenance. Cancer Cell. 9:425–434. 2006. View Article : Google Scholar
8	Zhao YH, Zhou M, Liu H, et al: Upregulation of lactate dehydrogenase A by ErbB2 through heat shock factor 1 promotes breast cancer cell glycolysis and growth. Oncogene. 28:3689–3701. 2009. View Article : Google Scholar : PubMed/NCBI
9	Le A, Cooper CR, Gouw AM, et al: Inhibition of lactate dehydrogenase A induces oxidative stress and inhibits tumor progression. Proc Natl Acad Sci USA. 107:2037–2042. 2010.PubMed/NCBI
10	Serganova I, Rizwan A, Ni XH, et al: Metabolic imaging: a link between lactate dehydrogenase A, lactate, and tumor phenotype. Clin Cancer Res. 17:6250–6261. 2011. View Article : Google Scholar : PubMed/NCBI
11	Wang ZY, Loo TY, Shen JG, et al: LDH-A silencing suppresses breast cancer tumorigenicity through induction of oxidative stress mediated mitochondrial pathway apoptosis. Breast Cancer Res Treat. 131:791–800. 2012. View Article : Google Scholar
12	Xie H, Valera VA, Merino MJ, et al: LDH-A inhibition, a therapeutic strategy for treatment of hereditary leiomyomatosis and renal cell cancer. Mol Cancer Ther. 8:626–635. 2009. View Article : Google Scholar : PubMed/NCBI
13	Sheng SL, Liu JJ, Dai YH, Sun XG, Xiong XP and Huang G: Knockdown of lactate dehydrogenase A suppresses tumor growth and metastasis of human hepatocellular carcinoma. FEBS J. 279:3898–3910. 2012. View Article : Google Scholar : PubMed/NCBI
14	Koukourakis MI, Giatromanolaki A, Winter S, Leek R, Sivridis E and Harris AL: Lactate dehydrogenase 5 expression in squamous cell head and neck cancer relates to prognosis following radical or postoperative radiotherapy. Oncology. 77:285–292. 2009. View Article : Google Scholar
15	Zhou GQ, Tang LL, Mao YP, et al: Baseline serum lactate dehydrogenase levels for patients treated with intensity-modulated radiotherapy for nasopharyngeal carcinoma: a predictor of poor prognosis and subsequent liver metastasis. Int J Radiat Oncol Biol Phys. 82:e359–e365. 2012. View Article : Google Scholar
16	Wan XB, Wei L, Li H, et al: High pretreatment serum lactate dehydrogenase level correlates with disease relapse and predicts an inferior outcome in locally advanced nasopharyngeal carcinoma. Eur J Cancer. 9:2356–2364. 2013. View Article : Google Scholar
17	Papaconstantinou J and Colowick SP: The role of glycolysis in the growth of tumor cells. II. The effect of oxamic acid on the growth of HeLa cells in tissue culture. J Biol Chem. 236:285–288. 1961.PubMed/NCBI
18	Thornburg JM, Nelson KK, Clem BF, et al: Targeting aspartate aminotransferase in breast cancer. Breast Cancer Res. 10:R842008. View Article : Google Scholar : PubMed/NCBI
19	Fiume L, Manerba M, Vettraino M and Di Stefano G: Impairment of aerobic glycolysis by inhibitors of lactic dehydrogenase hinders the growth of human hepatocellular carcinoma cell lines. Pharmacology. 86:157–162. 2010. View Article : Google Scholar : PubMed/NCBI
20	Zhou M, Zhao YH, Ding Y, et al: Warburg effect in chemosensitivity: targeting lactate dehydrogenase-A re-sensitizes Taxol-resistant cancer cells to Taxol. Mol Cancer. 9:332010. View Article : Google Scholar : PubMed/NCBI
21	Fiume L, Vettraino M, Manerba M and Di Stefano G: Inhibition of lactic dehydrogenase as a way to increase the anti-proliferative effect of multi-targeted kinase inhibitors. Pharmacol Res. 63:328–334. 2011. View Article : Google Scholar : PubMed/NCBI
22	Zhao YH, Liu H, Liu ZX, et al: Overcoming trastuzumab resistance in breast cancer by targeting dysregulated glucose metabolism. Cancer Res. 71:4585–4597. 2011. View Article : Google Scholar : PubMed/NCBI
23	Shapiro GI and Harper JW: Anticancer drug targets: cell cycle and checkpoint control. J Clin Invest. 104:1645–1653. 1999. View Article : Google Scholar : PubMed/NCBI
24	Simon HU, Haj-Yehia A and Levi-Schaffer F: Role of reactive oxygen species (ROS) in apoptosis induction. Apoptosis. 5:415–418. 2000.PubMed/NCBI
25	Modica-Napolitano JS and Singh KK: Mitochondrial dysfunction in cancer. Mitochondrion. 4:755–762. 2004.
26	Pelicano H, Lu W, Zhou Y, et al: Mitochondrial dysfunction and reactive oxygen species imbalance promote breast cancer cell motility through a CXCL14-mediated mechanism. Cancer Res. 69:2375–2383. 2009. View Article : Google Scholar
27	Hung WY, Huang KH, Wu CW, et al: Mitochondrial dysfunction promotes cell migration via reactive oxygen species-enhanced β5-integrin expression in human gastric cancer SC-M1 cells. Biochim Biophys Acta. 1820:1102–1110. 2012.PubMed/NCBI
28	Lindqvist A, van Zon W, Karlsson Rosenthal C and Wolthuis RM: Cyclin B1-Cdk1 activation continues after centrosome separation to control mitotic progression. PLoS Biol. 5:e1232007. View Article : Google Scholar : PubMed/NCBI
29	Madhok BM, Yeluri S, Perry SL, Hughes TA and Jayne DG: Dichloroacetate induces apoptosis and cell-cycle arrest in colorectal cancer cells. Br J Cancer. 102:1746–1752. 2010. View Article : Google Scholar : PubMed/NCBI
30	Leach JK, Van Tuyle G, Lin PS, Schmidt-Ullrich R and Mikkelsen RB: Ionizing radiation-induced, mitochondria-dependent generation of reactive oxygen/nitrogen. Cancer Res. 61:3894–3901. 2001.PubMed/NCBI
31	Tominaga H, Kodama S, Matsuda N, Suzuki K and Watanabe M: Involvement of reactive oxygen species (ROS) in the induction of genetic instability by radiation. J Radiat Res. 45:181–188. 2004. View Article : Google Scholar : PubMed/NCBI
32	Diehn M, Cho RW, Lobo NA, et al: Association of reactive oxygen species levels and radioresistance in cancer stem cells. Nature. 458:780–783. 2009. View Article : Google Scholar : PubMed/NCBI
33	Cao WG, Yacoub S, Shiverick KT, et al: Dichloroacetate (DCA) sensitizes both wild-type and overexpressing Bcl-2 prostate cancer cells in vitro to radiation. Prostate. 68:1223–1231. 2008. View Article : Google Scholar : PubMed/NCBI
34	Sharma PK, Bhardwaj R, Dwarakanath BS and Varshney R: Metabolic oxidative stress induced by a combination of 2-DG and 6-AN enhances radiation damage selectively in malignant cells via non-coordinated expression of antioxidant enzymes. Cancer Lett. 295:154–166. 2010. View Article : Google Scholar
35	Sandulache VC, Skinner HD, Wang Y, et al: Glycolytic inhibition alters anaplastic thyroid carcinoma tumor metabolism and improves response to conventional chemotherapy and radiation. Mol Cancer Ther. 11:1373–1380. 2012. View Article : Google Scholar
36	Goldberg EB and Colowick SP: The role of glycolysis in the growth of tumor cells. 3. Lactic dehydrogenase as the site of action of oxamate on the growth of cultured cells. J Biol Chem. 240:2786–2790. 1965.PubMed/NCBI
37	Elwood JC: Effect of oxamate on glycolysis and respiration in sarcoma 37 ascites cells. Cancer Res. 28:2056–2060. 1968.PubMed/NCBI
38	Granchi C, Bertini S, Macchia M and Minutolo F: Inhibitors of lactate dehydrogenase isoforms and their therapeutic potentials. Curr Med Chem. 17:672–697. 2010. View Article : Google Scholar : PubMed/NCBI
39	Granchi C, Roy S, Giacomelli C, et al: Discovery of N-hydroxyindole-based inhibitors of human lactate dehydrogenase isoform A (LDH-A) as starvation agents against cancer cells. J Med Chem. 54:1599–1612. 2011. View Article : Google Scholar
40	Wang Z, Wang N, Chen J and Shen J: Emerging glycolysis targeting and drug discovery from Chinese medicine in cancer therapy. Evid Based Complement Alternat Med. 2012:8731752012.PubMed/NCBI