Role of malate dehydrogenase in facilitating lactate dehydrogenase to support the glycolysis pathway in tumors

Mansouri,Siavash; Shahriari,Ali; Kalantar,Hadi; Moini Zanjani,Taraneh; Haghi Karamallah,Mojtaba

doi:10.3892/br.2017.873

Role of malate dehydrogenase in facilitating lactate dehydrogenase to support the glycolysis pathway in tumors

Authors:
- Siavash Mansouri
- Ali Shahriari
- Hadi Kalantar
- Taraneh Moini Zanjani
- Mojtaba Haghi Karamallah
View Affiliations

Affiliations: Department of Biochemistry and Molecular Biology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Khuzestan 61357‑83151, Iran, Department of Pharmacology, School of Pharmacy, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Khuzestan 61357-33184, Iran, Department of Pharmacology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran 19857‑17443, Iran, Department of Biochemistry and Molecular Biology, International Campus, Shahid Sadoughi University of Medical Sciences, Yazd 89151‑73143, Iran
Published online on: March 14, 2017 https://doi.org/10.3892/br.2017.873
Pages: 463-467

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

High aerobic glycolysis, as one of the hallmarks of cancer cells, requires nicotinamide adenine dinucleotide (NAD+) as a vital co‑factor, to guarantee the flow of glycolysis. Malate dehydrogenase (MDH), as an important enzyme in cancer metabolism, is a source of NAD+ additional to lactate dehydrogenase (LDH). The current study aimed to elucidate the kinetic parameters of MDH in human breast cancer and evaluate its supportive role in the glycolysis pathway. The Michaelis‑Menten constant (Km) and maximum velocity (Vmax) of MDH were determined in the crude extracts of human breast tumors and healthy tissue samples, which were obtained directly from the operating theatre. To assess the potential role of MDH in supporting glycolysis, the MDH activity was measured when the LDH activity was inhibited by different concentrations of oxamate, an inhibitor of LDH in breast cancer cell lines. The Km of cancerous MDH (C‑MDH) was the same as the healthy MDH, although the Vmax of C‑MDH was higher relative to the healthy MDH. Notably, the MDH activity was increased in the MDA‑MB‑231 cell line, which was treated with the LDH inhibitor (oxamate), but not in the MCF‑7 cell line (P<0.05). The higher tendency of C‑MDH for NAD+ and malate generation in cancer cells is an effective approach for supporting glycolysis. Increasing MDH activity in the absence of LDH demonstrates the supportive role of MDH in glycolysis. Therefore, decreasing MDH activity and expression in a forward reaction may present as a valid molecular target to abolish its potential effect on tumor metabolism.

View Figures

View References

1	Hanahan D and Weinberg RA: Hallmarks of cancer: The next generation. Cell. 144:646–674. 2011. View Article : Google Scholar : PubMed/NCBI
2	Warburg O: On the origin of cancer cells. Science. 123:309–314. 1956. View Article : Google Scholar : PubMed/NCBI
3	Gatenby RA and Gillies RJ: Why do cancers have high aerobic glycolysis? Nat Rev Cancer. 4:891–899. 2004. View Article : Google Scholar : PubMed/NCBI
4	Talaiezadeh A, Shahriari A, Tabandeh MR, Fathizadeh P and Mansouri S: Kinetic characterization of lactate dehydrogenase in normal and malignant human breast tissues. Cancer Cell Int. 15:192015. View Article : Google Scholar : PubMed/NCBI
5	Koukourakis MI, Giatromanolaki A, Simopoulos C, Polychronidis A and Sivridis E: Lactate dehydrogenase 5 (LDH5) relates to up-regulated hypoxia inducible factor pathway and metastasis in colorectal cancer. Clin Exp Metastasis. 22:25–30. 2005. View Article : Google Scholar : PubMed/NCBI
6	Rong Y, Wu W, Ni X, Kuang T, Jin D, Wang D and Lou W: Lactate dehydrogenase A is overexpressed in pancreatic cancer and promotes the growth of pancreatic cancer cells. Tumour Biol. 34:1523–1530. 2013. View Article : Google Scholar : PubMed/NCBI
7	Israël M and Schwartz L: The metabolic advantage of tumor cells. Mol Cancer. 10:702011. View Article : Google Scholar : PubMed/NCBI
8	DeBerardinis RJ, Lum JJ, Hatzivassiliou G and Thompson CB: The biology of cancer: Metabolic reprogramming fuels cell growth and proliferation. Cell Metab. 7:11–20. 2008. View Article : Google Scholar : PubMed/NCBI
9	Murray RK, Granner DK, Mayes PA and Rodwell VW: Harper's illustrated biochemistry. 26th edition. McGraw-Hill; New York, NY: 2003
10	Storey KB and Brooks SPJ: The basis of enzymatic adaptation. Principles of Medical Biology. 4:Part 1147–169. 1995. View Article : Google Scholar
11	Fukumura D and Jain RK: Tumor microenvironment abnormalities: Causes, consequences, and strategies to normalize. J Cell Biochem. 101:937–949. 2007. View Article : Google Scholar : PubMed/NCBI
12	Cairns RA, Harris IS and Mak TW: Regulation of cancer cell metabolism. Nat Rev Cancer. 11:85–95. 2011. View Article : Google Scholar : PubMed/NCBI
13	Grisham MB, Bernstein LH and Everse J: The cytoplasmic malate dehydrogenase in neoplastic tissues; presence of a novel isoenzyme? Br J Cancer. 47:727–731. 1983. View Article : Google Scholar : PubMed/NCBI
14	Belfiore F, Borzi V, Vecchio LL, Napoli E and Rabuazzo AM: Enzyme activities of NADPH-forming metabolic pathways in normal and leukemic leukocytes. Clin Chem. 21:880–883. 1975.PubMed/NCBI
15	Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 72:248–254. 1976. View Article : Google Scholar : PubMed/NCBI
16	Brooks SPJA: A simple computer program with statistical tests for the analysis of enzyme kinetics. Biotechniques. 13:906–911. 1992.PubMed/NCBI
17	Brooks SPJA: A program for analyzing enzyme rate data obtained from a microplate reader. Biotechniques. 17:1154–1161. 1994.PubMed/NCBI
18	Koukourakis MI, Kontomanolis E, Giatromanolaki A, Sivridis E and Liberis V: Serum and tissue LDH levels in patients with breast/gynaecological cancer and benign diseases. Gynecol Obstet Invest. 67:162–168. 2009. View Article : Google Scholar : PubMed/NCBI
19	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 : PubMed/NCBI
20	Deberardinis RJ, Sayed N, Ditsworth D and Thompson CB: Brick by brick: Metabolism and tumor cell growth. Curr Opin Genet Dev. 18:54–61. 2008. View Article : Google Scholar : PubMed/NCBI
21	Robey IF, Lien AD, Welsh SJ, Baggett BK and Gillies RJ: Hypoxia-inducible factor-1α and the glycolytic phenotype in tumors. Neoplasia. 7:324–330. 2005. View Article : Google Scholar : PubMed/NCBI
22	Allison SJ, Knight JR, Granchi C, Rani R, Minutolo F, Milner J and Phillips RM: Identification of LDH-A as a therapeutic target for cancer cell killing via (i) p53/NAD(H)-dependent and (ii) p53-independent pathways. Oncogenesis. 3:e1022014. View Article : Google Scholar : PubMed/NCBI
23	Shi Y and Pinto BM: Human lactate dehydrogenase a inhibitors: A molecular dynamics investigation. PLoS One. 9:e863652014. View Article : Google Scholar : PubMed/NCBI
24	Balinsky D, Platz CE and Lewis JW: Isozyme patterns of normal, benign, and malignant human breast tissues. Cancer Res. 43:5895–5901. 1983.PubMed/NCBI