Pyruvate kinase type M2 contributes to the development of pancreatic ductal adenocarcinoma by regulating the production of metabolites and reactive oxygen species

Yokoyama,Misa; Tanuma,Nobuhiro; Shibuya,Rie; Shiroki,Takeharu; Abue,Makoto; Yamamoto,Kuniharu; Miura,Koh; Yamaguchi,Kazunori; Sato,Ikuro; Tamai,Keiichi; Satoh,Kennichi

doi:10.3892/ijo.2018.4258

Pyruvate kinase type M2 contributes to the development of pancreatic ductal adenocarcinoma by regulating the production of metabolites and reactive oxygen species

Authors:
- Misa Yokoyama
- Nobuhiro Tanuma
- Rie Shibuya
- Takeharu Shiroki
- Makoto Abue
- Kuniharu Yamamoto
- Koh Miura
- Kazunori Yamaguchi
- Ikuro Sato
- Keiichi Tamai
- Kennichi Satoh
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Affiliations: Division of Cancer Stem Cell, Miyagi Cancer Center Research Institute, Natori, Miyagi 981-1293, Japan, Division of Cancer Chemotherapy, Miyagi Cancer Center Research Institute, Natori, Miyagi 981-1293, Japan, Department of Gastroenterology, Miyagi Cancer Center, Natori, Miyagi 981-1293, Japan, Department of Gastroenterological Surgery, Miyagi Cancer Center, Natori, Miyagi 981-1293, Japan, Division of Molecular and Cellular Oncology, Miyagi Cancer Center Research Institute, Natori, Miyagi 981-1293, Japan, Department of Pathology, Miyagi Cancer Center, Natori, Miyagi 981-1293, Japan
Published online on: January 30, 2018 https://doi.org/10.3892/ijo.2018.4258
Pages: 881-891

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Abstract

The majority of cancer cells maintain a high glycolytic activity and an increased lactate production, even in a well oxygenated environment. This phenomenon is known as the Warburg effect. Previous studies have revealed that various types of cancer selectively express the pyruvate kinase M2 isoform (PKM2), and that PKM2 plays a pivotal role in the Warburg effect. Although elevated PKM2 levels have been observed in pancreatic cancer and other types of cancer, little is known about the biological function of PKM2. In this study, in order to examine the expression and role of PKM2 in pancreatic ductal adenocarcinoma (PDAC), we knocked down PKM2 in PDAC cells by introducing small interfering and short hairpin RNAs, and examined the gene expression profiles in the cells by microarray analysis. We analyzed the energy-producing pathways in the cells by XFe Extracellular Flux Analyzers, and detected intracellular metabolites by capillary electrophoresis time-of-flight mass spectrometry. We found that the RNAi-mediated knockdown of PKM2 diminished the proliferative, migratory and tumorigenic ability of the PDAC cell-lines. PKM2 knockdown also resulted in lower glycolytic activities and decreased levels of some intracellular metabolites, such as pyruvate and polyamine; however, it led to elevated levels of reactive oxygen species. Microarray analysis revealed the functional association between PKM2 and the expression of genes that drive the cell cycle. On the whole, the findings of this study demonstrate that PKM2 plays an important role in metabolic activities, as well as in the malignancy of PDAC cells.

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1	Ma J, Siegel R and Jemal A: Pancreatic cancer death rates by race among US men and women, 1970–2009. J Natl Cancer Inst. 105:1694–1700. 2013. View Article : Google Scholar : PubMed/NCBI
2	Vander Heiden MG, Cantley LC and Thompson CB: Understanding the Warburg effect: The metabolic requirements of cell proliferation. Science. 324:1029–1033. 2009. View Article : Google Scholar : PubMed/NCBI
3	Christofk HR, Vander Heiden MG, Harris MH, Ramanathan A, Gerszten RE, Wei R, Fleming MD, Schreiber SL and Cantley LC: The M2 splice isoform of pyruvate kinase is important for cancer metabolism and tumour growth. Nature. 452:230–233. 2008. View Article : Google Scholar : PubMed/NCBI
4	Mazurek S: Pyruvate kinase type M2: A key regulator of the metabolic budget system in tumor cells. Int J Biochem Cell Biol. 43:969–980. 2011. View Article : Google Scholar
5	Marie J, Levin MJ, Simon MP and Kahn A: Genetic and epigenetic control of the pyruvate kinase isozymes in mammals. Isozymes Curr Top Biol Med Res. 7:221–240. 1983.PubMed/NCBI
6	Dong G, Mao Q, Xia W, Xu Y, Wang J, Xu L and Jiang F: PKM2 and cancer: The function of PKM2 beyond glycolysis. Oncol Lett. 11:1980–1986. 2016. View Article : Google Scholar : PubMed/NCBI
7	Li C, Zhao Z, Zhou Z and Liu R: PKM2 promotes cell survival and invasion under metabolic stress by enhancing Warburg effect in pancreatic ductal adenocarcinoma. Dig Dis Sci. 61:767–773. 2016. View Article : Google Scholar
8	Yang W, Xia Y, Cao Y, Zheng Y, Bu W, Zhang L, You MJ, Koh MY, Cote G, Aldape K, et al: EGFR-induced and PKCε monoubiquitylation-dependent NF-κB activation upregulates PKM2 expression and promotes tumorigenesis. Mol Cell. 48:771–784. 2012. View Article : Google Scholar : PubMed/NCBI
9	Wong N, Ojo D, Yan J and Tang D: PKM2 contributes to cancer metabolism. Cancer Lett. 356:184–191. 2015. View Article : Google Scholar
10	Luo W, Hu H, Chang R, Zhong J, Knabel M, O'Meally R, Cole RN, Pandey A and Semenza GL: Pyruvate kinase M2 is a PHD3-stimulated coactivator for hypoxia-inducible factor 1. Cell. 145:732–744. 2011. View Article : Google Scholar : PubMed/NCBI
11	Yang W, Xia Y, Hawke D, Li X, Liang J, Xing D, Aldape K, Hunter T, Alfred Yung WK and Lu Z: PKM2 phosphorylates histone H3 and promotes gene transcription and tumorigenesis. Cell. 150:685–696. 2012. View Article : Google Scholar : PubMed/NCBI
12	Goldberg MS and Sharp PA: Pyruvate kinase M2-specific siRNA induces apoptosis and tumor regression. J Exp Med. 209:217–224. 2012. View Article : Google Scholar : PubMed/NCBI
13	Bell SP and Dutta A: DNA replication in eukaryotic cells. Annu Rev Biochem. 71:333–374. 2002. View Article : Google Scholar : PubMed/NCBI
14	Pegg AE: Spermidine/spermine-N(1)-acetyltransferase: A key metabolic regulator. Am J Physiol Endocrinol Metab. 294:E995–E1010. 2008. View Article : Google Scholar : PubMed/NCBI
15	Lockney NA, Zhang M, Lu Y, Sopha SC, Washington MK, Merchant N, Zhao Z, Shyr Y, Chakravarthy AB and Xia F: Pyruvate kinase muscle isoenzyme 2 (PKM2) expression is associated with overall survival in pancreatic ductal adenocarcinoma. J Gastrointest Cancer. 46:390–398. 2015. View Article : Google Scholar : PubMed/NCBI
16	Pegg AE: Polyamine metabolism and its importance in neoplastic growth and a target for chemotherapy. Cancer Res. 48:759–774. 1988.PubMed/NCBI
17	Pegg AE: Mammalian polyamine metabolism and function. IUBMB Life. 61:880–894. 2009. View Article : Google Scholar : PubMed/NCBI
18	Childs AC, Mehta DJ and Gerner EW: Polyamine-dependent gene expression. Cell Mol Life Sci. 60:1394–1406. 2003. View Article : Google Scholar : PubMed/NCBI
19	Nowotarski SL, Woster PM and Casero RA Jr: Polyamines and cancer: Implications for chemotherapy and chemoprevention. Expert Rev Mol Med. 15:e32013. View Article : Google Scholar : PubMed/NCBI
20	Takigawa M, Verma AK, Simsiman RC and Boutwell RK: Inhibition of mouse skin tumor promotion and of promoter-stimulated epidermal polyamine biosynthesis by alpha-difluoromethylornithine. Cancer Res. 43:3732–3738. 1983.PubMed/NCBI
21	Gupta ED, Pachauri M, Ghosh PC and Rajam MV: Targeting polyamine biosynthetic pathway through RNAi causes the abrogation of MCF 7 breast cancer cell line. Tumour Biol. 37:1159–1171. 2016. View Article : Google Scholar
22	Russell DH: Increased polyamine concentrations in the urine of human cancer patients. Nat New Biol. 233:144–145. 1971. View Article : Google Scholar : PubMed/NCBI
23	Krüger A, Vowinckel J, Mülleder M, Grote P, Capuano F, Bluemlein K and Ralser M: Tpo1-mediated spermine and spermidine export controls cell cycle delay and times antioxidant protein expression during the oxidative stress response. EMBO Rep. 14:1113–1119. 2013. View Article : Google Scholar : PubMed/NCBI
24	Cross CE, Halliwell B, Borish ET, Pryor WA, Ames BN, Saul RL, McCord JM and Harman D: Oxygen radicals and human disease. Ann Intern Med. 107:526–545. 1987. View Article : Google Scholar : PubMed/NCBI
25	Chio IIC and Tuveson DA: ROS in cancer: The burning question. Trends Mol Med. 23:411–429. 2017. View Article : Google Scholar : PubMed/NCBI
26	Liou GY and Storz P: Reactive oxygen species in cancer. Free Radic Res. 44:479–496. 2010. View Article : Google Scholar : PubMed/NCBI
27	Galadari S, Rahman A, Pallichankandy S and Thayyullathil F: Reactive oxygen species and cancer paradox: To promote or to suppress? Free Radic Biol Med. 104:144–164. 2017. View Article : Google Scholar : PubMed/NCBI
28	Diehn M, Cho RW, Lobo NA, Kalisky T, Dorie MJ, Kulp AN, Qian D, Lam JS, Ailles LE, Wong M, 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
29	Zheng B, Peng J, Mollayup A, Bakri A, Guo L, Zheng J and Xu H: Construction of a prognostic prediction system for pancreatic ductal adenocarcinoma to investigate the key prognostic genes. Mol Med Rep. 17:216–224. 2018.
30	Badea L, Herlea V, Dima SO, Dumitrascu T and Popescu I: Combined gene expression analysis of whole-tissue and microdissected pancreatic ductal adenocarcinoma identifies genes specifically overexpressed in tumor epithelia. Hepatogastroenterology. 55:2016–2027. 2008.