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Metabolic reprogramming by PCK1 promotes TCA cataplerosis, oxidative stress and apoptosis in liver cancer cells and suppresses hepatocellular carcinoma

Oncogene volume 37, pages 1637–1653 (2018)Cite this article

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Abstract

Phosphoenolpyruvate carboxykinase (PEPCK or PCK) catalyzes the first rate-limiting step in hepatic gluconeogenesis pathway to maintain blood glucose levels. Mammalian cells express two PCK genes, encoding for a cytoplasmic (PCPEK-C or PCK1) and a mitochondrial (PEPCK-M or PCK2) isoforms, respectively. Increased expressions of both PCK genes are found in cancer of several organs, including colon, lung, and skin, and linked to increased anabolic metabolism and cell proliferation. Here, we report that the expressions of both PCK1 and PCK2 genes are downregulated in primary hepatocellular carcinoma (HCC) and low PCK expression was associated with poor prognosis in patients with HCC. Forced expression of either PCK1 or PCK2 in liver cancer cell lines results in severe apoptosis under the condition of glucose deprivation and suppressed liver tumorigenesis in mice. Mechanistically, we show that the pro-apoptotic effect of PCK1 requires its catalytic activity. We demonstrate that forced PCK1 expression in glucose-starved liver cancer cells induced TCA cataplerosis, leading to energy crisis and oxidative stress. Replenishing TCA intermediate α-ketoglutarate or inhibition of reactive oxygen species production blocked the cell death caused by PCK expression. Taken together, our data reveal that PCK1 is detrimental to malignant hepatocytes and suggest activating PCK1 expression as a potential treatment strategy for patients with HCC.

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References

  1. Hsu PP, Sabatini DM. Cancer cell metabolism: Warburg and beyond. Cell. 2008;134:703–7.

    Article  CAS  Google Scholar 

  2. Vander Heiden MG, Cantley LC, Thompson CB. Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science. 2009;324:1029–33.

    Article  Google Scholar 

  3. Warburg O. On the origin of cancer cells. Science. 1956;123:309–14.

    Article  CAS  Google Scholar 

  4. Burgess SC, He T, Yan Z, Lindner J, Sherry AD, Malloy CR, et al. Cytosolic phosphoenolpyruvate carboxykinase does not solely control the rate of hepatic gluconeogenesis in the intact mouse liver. Cell Metab. 2007;5:313–20.

    Article  CAS  Google Scholar 

  5. Yang J, Kalhan SC, Hanson RW. What is the metabolic role of phosphoenolpyruvate carboxykinase? J Biol Chem. 2009;284:27025–9.

    Article  CAS  Google Scholar 

  6. Méndez-Lucas A, Duarte JA, Sunny NE, Satapati S, He T, Fu X, et al. PEPCK-M expression in mouse liver potentiates, not replaces, PEPCK-C mediated gluconeogenesis. J Hepatol. 2013;59:105–13.

    Article  Google Scholar 

  7. Montal ED, Dewi R, Bhalla K, Ou L, Hwang BJ, Ropell AE, et al. PEPCK coordinates the regulation of central carbon metabolism to promote cancer cell growth. Mol Cell. 2015;60:571–83.

    Article  CAS  Google Scholar 

  8. Vincent EE, Sergushichev A, Griss T, Gingras MC, Samborska B, Ntimbane T, et al. Mitochondrial phosphoenolpyruvate carboxykinase regulates metabolic adaptation and enables glucose-independent tumor growth. Mol Cell. 2015;60:195–207.

    Article  CAS  Google Scholar 

  9. Leithner K, Hrzenjak A, Trötzmüller M, Moustafa T, Köfeler HC, Wohlkoenig C, et al. PCK2 activation mediates an adaptive response to glucose depletion in lung cancer. Oncogene. 2014;34:1044–50.

    Article  Google Scholar 

  10. Sunny NE, Parks EJ, Browning JD, Burgess SC. Excessive hepatic mitochondrial TCA cycle and gluconeogenesis in humans with nonalcoholic fatty liver disease. Cell Metab. 2011;14:804–10.

    Article  CAS  Google Scholar 

  11. Satapati S, Kucejova B, Duarte JA, Fletcher JA, Reynolds L, Sunny NE, et al. Mitochondrial metabolism mediates oxidative stress and inflammation in fatty liver. J Clin Invest. 2015;125:4447–62.

    Article  Google Scholar 

  12. Owen OE, Kalhan SC, Hanson RW. The key role of anaplerosis and cataplerosis for citric acid cycle function. J Biol Chem. 2002;277:30409–12.

    Article  CAS  Google Scholar 

  13. Beale EG, Harvey BJ, Forest C. PCK1 and PCK2 as candidate diabetes and obesity genes. Cell Biochem Biophys. 2007;48:89–95.

    Article  CAS  Google Scholar 

  14. Hirayama A, Kami K, Sugimoto M, Sugawara M, Toki N, Onozuka H, et al. Quantitative metabolome profiling of colon and stomach cancer microenvironment by capillary electrophoresis time-of-flight mass spectrometry. Cancer Res. 2009;69:4918–25.

    Article  CAS  Google Scholar 

  15. Dunten P, Belunis C, Crowther R, Hollfelder K, Kammlott U, Levin W, et al. Crystal structure of human cytosolic phosphoenolpyruvate carboxykinase reveals a new GTP-binding site. J Mol Biol. 2001;316:257–64.

    Article  Google Scholar 

  16. Balan MD, McLeod MJ, Lotosky WR, Ghaly M, Holyoak T. Inhibition and allosteric regulation of monomeric phosphoenolpyruvate carboxykinase by 3-mercaptopicolinic acid. Biochemistry. 2015;54:5878–87.

    Article  CAS  Google Scholar 

  17. Burgess SC, Hausler N, Merritt M, Jeffrey FM, Storey C, Milde A, et al. Impaired tricarboxylic acid cycle activity in mouse livers lacking cytosolic phosphoenolpyruvate carboxykinase. J Biol Chem. 2004;279:48941–9.

    Article  CAS  Google Scholar 

  18. Hakimi P, Johnson MT, Yang J, Lepage DF, Conlon RA, Kalhan SC, et al. Phosphoenolpyruvate carboxykinase and the critical role of cataplerosis in the control of hepatic metabolism. Nutr Metab. 2005;2:33.

    Article  Google Scholar 

  19. Hitosugi T, Zhou L, Elf S, Fan J, Kang HB, Seo JH, et al. Phosphoglycerate mutase 1 coordinates glycolysis and biosynthesis to promote tumor growth. Cancer Cell. 2012;22:585–600.

    Article  CAS  Google Scholar 

  20. Sullivan LB, Martinez-Garcia E, Nguyen H, Mullen AR, Dufour E, Sudarshan S, et al. The proto-oncometabolite fumarate binds glutathione to amplify ROS-dependent signaling. Mol Cell. 2013;51:236–48.

    Article  CAS  Google Scholar 

  21. Zheng L, Cardaci S, Jerby L, MacKenzie ED, Sciacovelli M, Johnson TI, et al. Fumarate induces redox-dependent senescence by modifying glutathione metabolism. Nat Commun. 2015;6:6001.

    Article  CAS  Google Scholar 

  22. Jin L, Li D, Alesi GN, Fan J, Kang HB, Lu Z, et al. Glutamate dehydrogenase 1 signals through antioxidant glutathione peroxidase 1 to regulate redox homeostasis and tumor growth. Cancer Cell. 2015;27:257–70.

    Article  CAS  Google Scholar 

  23. Sabharwal SS, Schumacker PT. Mitochondrial ROS in cancer: initiators, amplifiers or an Achilles’ heel? Nat Rev Cancer. 2014;14:709–21.

    Article  CAS  Google Scholar 

  24. Murphy MP. How mitochondria produce reactive oxygen species. Biochem J. 2009;417:1–13.

    Article  CAS  Google Scholar 

  25. Shen S, Guo X, Yan H, Lu Y, Ji X, Li L, et al. A miR-130a-YAP positive feedback loop promotes organ size and tumorigenesis. Cell Res. 2015;25:997–1012.

    Article  CAS  Google Scholar 

  26. Hernandez-Gea V, Toffanin S, Friedman SL, Llovet JM. Role of the microenvironment in the pathogenesis and treatment of hepatocellular carcinoma. Gastroenterology. 2013;144:512–27.

    Article  Google Scholar 

  27. Tlsty TD, Coussens LM. Tumor stroma and regulation of cancer development. Annu Rev Pathol. 2006;1:119–50.

    Article  CAS  Google Scholar 

  28. Mao Y, Keller ET, Garfield DH, Shen K, Wang J. Stromal cells in tumor microenvironment and breast cancer. Cancer Metastas- Rev. 2013;32:303–15.

    Article  Google Scholar 

  29. Li B, Qiu B, Lee DS, Walton ZE, Ochocki JD, Mathew LK, et al. Fructose-1,6-bisphosphatase opposes renal carcinoma progression. Nature. 2014;513:251–5.

    Article  CAS  Google Scholar 

  30. Yang H, Zhou L, Shi Q, Zhao Y, Lin H, Zhang M, et al. SIRT3-dependent GOT2 acetylation status affects the malate-aspartate NADH shuttle activity and pancreatic tumor growth. EMBO J. 2015;34:1110–25.

    Article  CAS  Google Scholar 

  31. Ma S, Jiang B, Deng W, Gu ZK, Wu FZ, Li T, et al. D-2-hydroxyglutarate is essential for maintaining oncogenic property of mutant IDH-containing cancer cells but dispensable for cell growth. Oncotarget. 2015;6:8606–20.

    PubMed  PubMed Central  Google Scholar 

  32. Haslene-Hox H, Oveland E, Berg KC, Kolmannskog O, Woie K, Salvesen HB, et al. A new method for isolation of interstitial fluid from human solid tumors applied to proteomic analysis of ovarian carcinoma tissue. PLoS ONE. 2011;6:e19217.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Dr Bin Zhao (Zhejiang University, Hangzhou, China) for providing plasmids used for inducing liver tumor in mice, including PB[CMV-myc-YAP-5SA]DS, PB[Act-RFP]DS, and Act-PB Transposase. We thank Xiaocan Guo for technical help on hydrodynamic injection. We also thank Dr Jiahuai Han (Xiamen University, Xiamen, China) for generously offering cDNAs for human PCK1 and PCK2 genes. This work was supported by the National Science and Technology Major Project of China (No. 2017ZX10203207-002-004 to HXY), the 973 Program (No. 2015CB910401 to YX), the NSFC grants (No. 31570784 and 81773190 to HXY), and NIH grants (CA163834 to YX; CA196878 and GM51586 to K-LG).

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Authors and Affiliations

  1. The Molecular and Cell Biology Research Lab, Institute of Biomedical Sciences, Shanghai Medical College, Fudan University, Shanghai, China

    Meng-Xi Liu, Si-Jia Sun, Peng Liu, Xu Feng, Zhou-Li Cheng, Kun-Liang Guan, Hai-Xin Yuan & Yue Xiong

  2. Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, and Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China

    Meng-Xi Liu, Si-Jia Sun, Peng Liu, Xu Feng, Zhou-Li Cheng, Kun-Liang Guan, Hai-Xin Yuan & Yue Xiong

  3. State Key Laboratory of Genetic Engineering, Collaborative Innovation Center of Genetics and Development, School of Life Sciences, Fudan University, Shanghai, China

    Meng-Xi Liu, Kun-Liang Guan, Hai-Xin Yuan & Yue Xiong

  4. Department of Liver Surgery, Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai, China

    Lei Jin, Wei-Ren Liu & Ying-Hong Shi

  5. Key Laboratory of Carcinogenesis and Cancer Invasion of Ministry of Education, Shanghai, China

    Lei Jin, Wei-Ren Liu & Ying-Hong Shi

  6. Department of Pharmacology and Moores Cancer Center, University of California San Diego, La Jolla, CA, USA

    Kun-Liang Guan

  7. Department of Biochemistry and Biophysics, Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    Yue Xiong

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  1. Meng-Xi Liu
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Correspondence to Ying-Hong Shi, Hai-Xin Yuan or Yue Xiong.

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K-LG is co-founder of Vivace Therapeutics. The remaining authors declare that they have no competing interests.

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Liu, MX., Jin, L., Sun, SJ. et al. Metabolic reprogramming by PCK1 promotes TCA cataplerosis, oxidative stress and apoptosis in liver cancer cells and suppresses hepatocellular carcinoma. Oncogene 37, 1637–1653 (2018). https://doi.org/10.1038/s41388-017-0070-6

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