ETS-1: A potential target of glycolysis for metabolic therapy by regulating glucose metabolism in pancreatic cancer

Zhang,Xiu; Wu,Dan; Aldarouish,Mohanad; Yin,Xiaodong; Li,Chunyan; Wang,Cailian

doi:10.3892/ijo.2016.3770

ETS-1: A potential target of glycolysis for metabolic therapy by regulating glucose metabolism in pancreatic cancer

Authors:
- Xiu Zhang
- Dan Wu
- Mohanad Aldarouish
- Xiaodong Yin
- Chunyan Li
- Cailian Wang
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Affiliations: Department of Oncology, Zhongda Hospital, Medical School of Southeast University, Nanjing, Jiangsu 210009, P.R. China, Department of Oncology, Jiangyin People's Hospital, Jiangyin, Jiangsu 224000, P.R. China, Department of Oncology, Binhai People's Hospital, Yancheng, Jiangsu 224500, P.R. China, Nanjing Medical University, Nanjing, Jiangsu 210009, P.R. China
Published online on: November 16, 2016 https://doi.org/10.3892/ijo.2016.3770
Pages: 232-240

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Abstract

Pancreatic cancer is one of the most lethal malignancies of all types of cancer due to lack of early symptoms and its resistance to conventional therapy. In our previous study, we have shown that v‑ets erythroblastosis virus E26 oncogene homolog‑1 (ETS‑1) promote cell migration and invasion in pancreatic cancer cells. However, the function of ETS‑1 in regulation of glycolysis and autophagy during progression of pancreatic cancer has not been defined yet. In this study, we sought to identify the potential role for silencing ETS‑1 in reducing the expression of glucose transporter‑1 (GLUT‑1) to disturb glycolysis through alteration of ‘Warburg effect’, by which could result in AMP‑activated protein kinase (AMPK) activation, autophagy induction and reduction of cell viability. MTT assay was applied to assess the cell viability in ETS‑1 silencing cells and control groups. Glucose absorption rate, lactate production rate and cellular ATP level were measured by standard colorimetric assay kits. The levels of mRNAs of ETS‑1, GLUT‑1, autophagy‑related gene 5 (ATG5) and ATG7 were analyzed by qRT‑PCR. The expression of ETS‑1, GLUT‑1, ATG5, ATG7, p‑AMPK, and LC3II proteins were evaluated by western blot analysis. GraphPad Prism 5.0 was used for all statistical analysis. We found that cell viability was obviously attenuated after silencing ETS‑1. Besides, our results also showed that the expression of GLUT‑1 significantly declined in ETS‑1 silencing cell lines which resulted in a lower glucose utilization and lactate production. Furthermore, the inhibition of glycolysis, which depends on glucose utilization and lactate production, reduced the generation of energy in the form of ATP. Moreover, the reduction of cellular ATP was associated with stimulation of AMP‑activated protein kinase (AMPK) and induction of autophagy. Our results indicated that ETS‑1 induced autophagy after inhibition of glycolysis, and thus led to comparative decrease of cell viability. These results implied that ETS‑1 could be a potential target for tumor metabolic therapy.

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1	Perera RM and Bardeesy N: Pancreatic cancer metabolism: Breaking it down to build it back up. Cancer Discov. 5:1247–1261. 2015. View Article : Google Scholar : PubMed/NCBI
2	Xie D and Xie K: Pancreatic cancer stromal biology and therapy. Genes Dis. 2:133–143. 2015. View Article : Google Scholar : PubMed/NCBI
3	Cid-Arregui A and Juarez V: Perspectives in the treatment of pancreatic adenocarcinoma. World J Gastroenterol. 21:9297–9316. 2015. View Article : Google Scholar : PubMed/NCBI
4	Guillaumond F, Iovanna JL and Vasseur S: Pancreatic tumor cell metabolism: Focus on glycolysis and its connected metabolic pathways. Arch Biochem Biophys. 545:69–73. 2014. View Article : Google Scholar : PubMed/NCBI
5	Paniccia A, Merkow J, Edil BH and Zhu Y: Immunotherapy for pancreatic ductal adenocarcinoma: An overview of clinical trials. Chin J Cancer Res. 27:376–391. 2015.PubMed/NCBI
6	Tennant DA, Durán RV and Gottlieb E: Targeting metabolic transformation for cancer therapy. Nat Rev Cancer. 10:267–277. 2010. View Article : Google Scholar : PubMed/NCBI
7	Lee M and Yoon JH: Metabolic interplay between glycolysis and mitochondrial oxidation: The reverse Warburg effect and its therapeutic implication. World J Biol Chem. 6:148–161. 2015. View Article : Google Scholar : PubMed/NCBI
8	Zhou W, Capello M, Fredolini C, Piemonti L, Liotta LA, Novelli F and Petricoin EF: Proteomic analysis of pancreatic ductal adenocarcinoma cells reveals metabolic alterations. J Proteome Res. 10:1944–1952. 2011. View Article : Google Scholar : PubMed/NCBI
9	Lunt SY and Vander Heiden MG: Aerobic glycolysis: Meeting the metabolic requirements of cell proliferation. Annu Rev Cell Dev Biol. 27:441–464. 2011. View Article : Google Scholar : PubMed/NCBI
10	Li C, Wang Z, Chen Y, Zhou M, Zhang H, Chen R, Shi F, Wang C and Rui Z: Transcriptional silencing of ETS-1 abrogates epithelial-mesenchymal transition resulting in reduced motility of pancreatic cancer cells. Oncol Rep. 33:559–565. 2015.
11	Verschoor ML, Verschoor CP and Singh G: Ets-1 global gene expression profile reveals associations with metabolism and oxidative stress in ovarian and breast cancers. Cancer Metab. 1:172013. View Article : Google Scholar : PubMed/NCBI
12	Zhu M, Li M, Zhang F, Feng F, Chen W, Yang Y, Cui J, Zhang D and Linghu E: FBI-1 enhances ETS-1 signaling activity and promotes proliferation of human colorectal carcinoma cells. PLoS One. 9:e980412014. View Article : Google Scholar : PubMed/NCBI
13	Shaikhibrahim Z and Wernert N: ETS transcription factors and prostate cancer: The role of the family prototype ETS-1 (Review). Int J Oncol. 40:1748–1754. 2012.PubMed/NCBI
14	Lelièvre E, Lionneton F, Soncin F and Vandenbunder B: The Ets family contains transcriptional activators and repressors involved in angiogenesis. Int J Biochem Cell Biol. 33:391–407. 2001. View Article : Google Scholar : PubMed/NCBI
15	Oikawa T: ETS transcription factors: Possible targets for cancer therapy. Cancer Sci. 95:626–633. 2004. View Article : Google Scholar : PubMed/NCBI
16	Hahne JC, Okuducu AF, Kaminski A, Florin A, Soncin F and Wernert N: Ets-1 expression promotes epithelial cell transformation by inducing migration, invasion and anchorage-independent growth. Oncogene. 24:5384–5388. 2005. View Article : Google Scholar : PubMed/NCBI
17	Shaikhibrahim Z, Langer B, Lindstrot A, Florin A, Bosserhoff A, Buettner R and Wernert N: Ets-1 is implicated in the regulation of androgen co-regulator FHL2 and reveals specificity for migration, but not invasion, of PC3 prostate cancer cells. Oncol Rep. 25:1125–1129. 2011.PubMed/NCBI
18	Gao H, Peng C, Liang B, Shahbaz M, Liu S, Wang B, Sun Q, Niu Z, Niu W, Liu E, et al: β6 integrin induces the expression of metalloproteinase-3 and metalloproteinase-9 in colon cancer cells via ERK-ETS1 pathway. Cancer Lett. 354:427–437. 2014. View Article : Google Scholar : PubMed/NCBI
19	Bu S, Yamanaka M, Pei H, Bielawska A, Bielawski J, Hannun YA, Obeid L and Trojanowska M: Dihydrosphingosine 1-phosphate stimulates MMP1 gene expression via activation of ERK1/2-Ets1 pathway in human fibroblasts. FASEB J. 20:184–186. 2006.
20	Li T and Jiang S: Effect of bFGF on invasion of ovarian cancer cells through the regulation of Ets-1 and urokinase-type plasminogen activator. Pharm Biol. 48:161–165. 2010. View Article : Google Scholar : PubMed/NCBI
21	Jiang Y, Xu W, Lu J, He F and Yang X: Invasiveness of hepatocellular carcinoma cell lines: Contribution of hepatocyte growth factor, c-met, and transcription factor Ets-1. Biochem Biophys Res Commun. 286:1123–1130. 2001. View Article : Google Scholar : PubMed/NCBI
22	Tomita N, Morishita R, Taniyama Y, Koike H, Aoki M, Shimizu H, Matsumoto K, Nakamura T, Kaneda Y and Ogihara T: Angiogenic property of hepatocyte growth factor is dependent on upregulation of essential transcription factor for angiogenesis, ets-1. Circulation. 107:1411–1417. 2003. View Article : Google Scholar : PubMed/NCBI
23	Kato T, Fujita Y, Nakane K, Kojima T, Nozawa Y, Deguchi T and Ito M: ETS1 promotes chemoresistance and invasion of paclitaxel-resistant, hormone-refractory PC3 prostate cancer cells by upregulating MDR1 and MMP9 expression. Biochem Biophys Res Commun. 417:966–971. 2012. View Article : Google Scholar
24	Verschoor ML, Wilson LA, Verschoor CP and Singh G: Ets-1 regulates energy metabolism in cancer cells. PLoS One. 5:e135652010. View Article : Google Scholar : PubMed/NCBI
25	Sato T and Miwa A: Ets-1 and integrin beta3 for lung metastasis from colorectal cancer. APMIS. 110:347–353. 2002. View Article : Google Scholar : PubMed/NCBI
26	Sasaki H, Yukiue H, Moiriyama S, Kobayashi Y, Nakashima Y, Kaji M, Kiriyama M, Fukai I, Yamakawa Y and Fujii Y: Clinical significance of matrix metalloproteinase-7 and Ets-1 gene expression in patients with lung cancer. J Surg Res. 101:242–247. 2001. View Article : Google Scholar : PubMed/NCBI
27	Verschoor ML and Singh G: Ets-1 regulates intracellular glutathione levels: Key target for resistant ovarian cancer. Mol Cancer. 12:1382013. View Article : Google Scholar : PubMed/NCBI
28	Li W, Zang W, Liu P, Wang Y, Du Y, Chen X, Deng M, Sun W, Wang L, Zhao G, et al: MicroRNA-124 inhibits cellular proliferation and invasion by targeting Ets-1 in breast cancer. Tumour Biol. 35:10897–10904. 2014. View Article : Google Scholar : PubMed/NCBI
29	Tang N, Wang X, Huang T, Wu Y and Chen Y: Role of Ets-1 in fibronectin-derived heparin-binding domain polypeptides alleviating melanoma cell invasiveness and chemoresistance. Exp Dermatol. 23:512–513. 2014. View Article : Google Scholar : PubMed/NCBI
30	Span PN, Manders P, Heuvel JJ, Thomas CM, Bosch RR, Beex LV and Sweep CG: Expression of the transcription factor Ets-1 is an independent prognostic marker for relapse-free survival in breast cancer. Oncogene. 21:8506–8509. 2002. View Article : Google Scholar : PubMed/NCBI
31	Puzovic V, Brcic I, Ranogajec I and Jakic-Razumovic J: Prognostic values of ETS-1, MMP-2 and MMP-9 expression and co-expression in breast cancer patients. Neoplasma. 61:439–446. 2014. View Article : Google Scholar : PubMed/NCBI
32	Davidson B, Reich R, Goldberg I, Gotlieb WH, Kopolovic J, Berner A, Ben-Baruch G, Bryne M and Nesland JM: Ets-1 messenger RNA expression is a novel marker of poor survival in ovarian carcinoma. Clin Cancer Res. 7:551–557. 2001.PubMed/NCBI
33	Davidson B, Risberg B, Goldberg I, Nesland JM, Berner A, Tropé CG, Kristensen GB, Bryne M and Reich R: Ets-1 mRNA expression in effusions of serous ovarian carcinoma patients is a marker of poor outcome. Am J Surg Pathol. 25:1493–1500. 2001. View Article : Google Scholar : PubMed/NCBI
34	Takai N, Miyazaki T, Nishida M, Nasu K and Miyakawa I: c-Ets1 is a promising marker in epithelial ovarian cancer. Int J Mol Med. 9:287–292. 2002.PubMed/NCBI
35	Fujimoto J, Aoki I, Toyoki H, Khatun S and Tamaya T: Clinical implications of expression of ETS-1 related to angiogenesis in uterine cervical cancers. Ann Oncol. 13:1598–1604. 2002. View Article : Google Scholar : PubMed/NCBI
36	Kimmelman AC: Metabolic dependencies in RAS-driven cancers. Clin Cancer Res. 21:1828–1834. 2015. View Article : Google Scholar : PubMed/NCBI
37	Cairns RA: Drivers of the Warburg phenotype. Cancer J. 21:56–61. 2015. View Article : Google Scholar : PubMed/NCBI
38	Mogi A, Koga K, Aoki M, Hamasaki M, Uesugi N, Iwasaki A, Shirakusa T, Tamura K and Nabeshima K: Expression and role of GLUT-1, MCT-1, and MCT-4 in malignant pleural mesothelioma. Virchows Arch. 462:83–93. 2013. View Article : Google Scholar
39	Blum R and Kloog Y: Metabolism addiction in pancreatic cancer. Cell Death Dis. 5:e10652014. View Article : Google Scholar : PubMed/NCBI
40	Granja S, Pinheiro C, Reis RM, Martinho O and Baltazar F: Glucose addiction in cancer therapy: Advances and drawbacks. Curr Drug Metab. 16:221–242. 2015. View Article : Google Scholar : PubMed/NCBI
41	Li N, Huang D, Lu N and Luo L: Role of the LKB1/AMPK pathway in tumor invasion and metastasis of cancer cells (Review). Oncol Rep. 34:2821–2826. 2015.PubMed/NCBI
42	Kim N, Jeong S, Jing K, Shin S, Kim S, Heo JY, Kweon GR, Park SK, Wu T, Park JI, et al: Docosahexaenoic acid induces cell death in human non-small cell lung cancer cells by repressing mTOR via AMPK activation and PI3K/Akt inhibition. BioMed Res Int. 2015:2397642015. View Article : Google Scholar : PubMed/NCBI
43	Saito Y, Chapple RH, Lin A, Kitano A and Nakada D: AMPK protects leukemia-initiating cells in myeloid leukemias from metabolic stress in the bone marrow. Cell Stem Cell. 17:585–596. 2015. View Article : Google Scholar : PubMed/NCBI
44	Mihaylova MM and Shaw RJ: The AMPK signalling pathway coordinates cell growth, autophagy and metabolism. Nat Cell Biol. 13:1016–1023. 2011. View Article : Google Scholar : PubMed/NCBI
45	Kim J, Kundu M, Viollet B and Guan KL: AMPK and mTOR regulate autophagy through direct phosphorylation of Ulk1. Nat Cell Biol. 13:132–141. 2011. View Article : Google Scholar : PubMed/NCBI
46	Abdou AG, Eldien MM and Elsakka D: GLUT-1 Expression in cutaneous basal and squamous cell carcinomas. Int J Surg Pathol. 23:447–453. 2015. View Article : Google Scholar : PubMed/NCBI
47	Vuppalapati KK, Bouderlique T, Newton PT, Kaminskyy VO, Wehtje H, Ohlsson C, Zhivotovsky B and Chagin AS: Targeted deletion of autophagy genes Atg5 or Atg7 in the chondrocytes promotes caspase-dependent cell death and leads to mild growth retardation. J Bone Miner Res. 30:2249–2261. 2015. View Article : Google Scholar : PubMed/NCBI
48	Lao Y and Xu N: Autophagy in cancer chemoprevention: Identification of novel autophagy modulators with anticancer potential. Methods Mol Biol. 1379:151–163. 2016. View Article : Google Scholar
49	Shimizu S, Arakawa S and Nishida Y: Autophagy takes an alternative pathway. Autophagy. 6:290–291. 2010. View Article : Google Scholar : PubMed/NCBI
50	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
51	Zhang TB, Zhao Y, Tong ZX and Guan YF: Inhibition of glucose-transporter 1 (GLUT-1) expression reversed Warburg effect in gastric cancer cell MKN45. Int J Clin Exp Med. 8:2423–2428. 2015.PubMed/NCBI
52	Yoon SO, Jeon TJ, Park JS, Ryu YH, Lee JH, Yoo JS, Kim JK, Yoon DS and Oh EJ: Analysis of the roles of glucose transporter 1 and hexokinase 2 in the metabolism of glucose by extrahepatic bile duct cancer cells. Clin Nucl Med. 40:e178–e182. 2015. View Article : Google Scholar : PubMed/NCBI
53	Kim SE, Park HJ, Jeong HK, Kim MJ, Kim M, Bae ON and Baek SH: Autophagy sustains the survival of human pancreatic cancer PANC-1 cells under extreme nutrient deprivation conditions. Biochem Biophys Res Commun. 463:205–210. 2015. View Article : Google Scholar : PubMed/NCBI
54	Liu B, Cheng Y, Liu Q, Bao JK and Yang JM: Autophagic pathways as new targets for cancer drug development. Acta Pharmacol Sin. 31:1154–1164. 2010. View Article : Google Scholar : PubMed/NCBI