Skip to main content
SpringerLink
Log in

Malic enzyme 1 induces epithelial–mesenchymal transition and indicates poor prognosis in hepatocellular carcinoma

  • Research Article
  • Published:
Tumor Biology

Abstract

Malic enzyme 1 (ME1) links the glycolytic and citric acid cycles and is important for NADPH production, glutamine metabolism, and lipogenesis. Recently, its deregulation has been implicated in the progression of various cancers. However, the role of ME1 in the progression of hepatocellular carcinoma (HCC) remains unclear. In this study, we utilized short hairpin RNA-mediated gene silencing to investigate the biological effects of ME1 depletion in HCC and determined its prognostic significance in HCC. ME1 expression was examined by real-time (RT)-PCR and Western blot using five HCC cell lines and one normal liver cell line. We used polyethylenimine nanoparticles to deliver a short hairpin RNA to induce cessation of ME1 expression in HCC cells. Changes in NADPH production and reactive oxygen species (ROS) production were studied. Metastatic potentials of HCC cells were evaluated in vitro. Furthermore, we evaluated the protein level of ME1 in para-tumor and cancerous tissues of 65 HCC patients with detailed clinical, pathological, and clinical follow-up data. Patients’ survivals were further assessed as well. Upregulated ME1 expression was observed in HCC cell lines. Downregulation of ME1 attenuated NADPH production and stimulated ROS production. Silencing ME1 was noted to inhibit migratory and invasive properties of HCC cells by inducing the E-cadherin expression and decreasing of N-cadherin and vimentin expression in a ROS-dependent pathway. Overexpression of ME1 was observed in a major fraction of HCC samples. Higher level of ME1 in tumors was significantly associated with reduced overall survival (Kaplan–Meier analysis, P = 0.024) and reduced progression-free survival (Kaplan–Meier analysis, P = 0.011). Inhibition of ME1 expression decreases HCC metastasis via suppression of epithelial–mesenchymal transition (EMT) processes in ROS-induced pathways. ME1 overexpression associates with unfavorable prognoses in patients with HCC, suggesting that ME1 is a poor prognostic predictor of hepatocellular carcinoma.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Mazzanti R, Gramantieri L, Bolondi L. Hepatocellular carcinoma: epidemiology and clinical aspects. Mol Asp Med. 2008;29:130–43.

    Article  Google Scholar 

  2. Hu L, Xue F, Li Y, Shao M, Sun Y, Wei G. A long-term follow-up and comprehensive observation of risk and prognosis factors of recurrence and survival after resection of hepatocellular carcinoma. Cell Biochem Biophys. 2014;69:421–31. Springer US.

    Article  CAS  PubMed  Google Scholar 

  3. Kim J-W, Dang CV. Cancer’s molecular sweet tooth and the Warburg effect. Cancer Res. 2006;66:8927–30. American Association for Cancer Research.

    Article  CAS  PubMed  Google Scholar 

  4. Chen Z, Wang Z, Guo W, Zhang Z, Zhao F, Zhao Y, et al. TRIM35 interacts with pyruvate kinase isoform M2 to suppress the Warburg effect and tumorigenicity in hepatocellular carcinoma. Oncogene. 2014.

  5. Corominas-Faja B, Cuyàs E, Gumuzio J, Bosch-Barrera J, Leis O, Martin ÁG, et al. Chemical inhibition of acetyl-CoA carboxylase suppresses self-renewal growth of cancer stem cells. Oncotarget. 2014;5:8306–16.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Wang J, Yuan W, Chen Z, Wu S, Chen J, Ge J, et al. Overexpression of G6PD is associated with poor clinical outcome in gastric cancer. Tumour Biol. 2012;33:95–101. Springer Netherlands.

    Article  PubMed  Google Scholar 

  7. Jiang L, Xiao L, Sugiura H, Huang X, Ali A, Kuro-O M, et al. Metabolic reprogramming during TGFβ1-induced epithelial-to-mesenchymal transition. Oncogene. 2014;0.

  8. Jiang L, Wang H, Li J, Fang X, Pan H, Yuan X, et al. Up-regulated FASN expression promotes transcoelomic metastasis of ovarian cancer cell through epithelial-mesenchymal transition. Int J Mol Sci. 2014;15:11539–54.

    Article  PubMed  PubMed Central  Google Scholar 

  9. Wise EM, Ball EG. Malic enzyme and lipogenesis. Proc Natl Acad Sci U S A. 1964;52:1255–63.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Al-Dwairi A, Pabona JMP, Simmen RCM, Simmen FA. Cytosolic malic enzyme 1 (ME1) mediates high fat diet-induced adiposity, endocrine profile, and gastrointestinal tract proliferation-associated biomarkers in male mice. Singh SR, editor. PLoS ONE. Public Library of Science; 2012;7:e46716.

  11. Jiang P, Du W, Mancuso A, Wellen KE, Yang X. Reciprocal regulation of p53 and malic enzymes modulates metabolism and senescence. Nature. 2013;493:689–93.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Zheng F-J, Ye H-B, Wu M-S, Lian Y-F, Qian C-N, Zeng Y-X. Repressing malic enzyme 1 redirects glucose metabolism, unbalances the redox state, and attenuates migratory and invasive abilities in nasopharyngeal carcinoma cell lines. Chin J Cancer. 2012;31:519–31.

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Tang ZY, Sun FX, Tian J, Ye SL, Liu YK, Liu KD, et al. Metastatic human hepatocellular carcinoma models in nude mice and cell line with metastatic potential. World J Gastroenterol. 2001;7:597–601.

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Li Y, Tang ZY, Ye SL, Liu YK, Chen J, Xue Q, et al. Establishment of cell clones with different metastatic potential from the metastatic hepatocellular carcinoma cell line MHCC97. World J Gastroenterol. 2001;7:630–6.

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Pi H, Xu S, Zhang L, Guo P, Li Y, Xie J, et al. Dynamin 1-like-dependent mitochondrial fission initiates overactive mitophagy in the hepatotoxicity of cadmium. Autophagy. 2013;9:1780–800. Taylor & Francis.

    Article  CAS  PubMed  Google Scholar 

  16. Hu Y-J, Li H-Y, Qiu K-J, Li D-C, Zhou J-H, Hu Y-H, et al. Downregulation of Notch1 inhibits the invasion of human hepatocellular carcinoma HepG2 and MHCC97H cells through the regulation of PTEN and FAK. Int J Mol Med. 2014;34:1081–6. Spandidos Publications.

    CAS  PubMed  Google Scholar 

  17. Sun FX, Tang ZY, Lui KD, Ye SL, Xue Q, Gao DM, et al. Establishment of a metastatic model of human hepatocellular carcinoma in nude mice via orthotopic implantation of histologically intact tissues. Int J Cancer. 1996;66:239–43. Wiley Subscription Services, Inc., A Wiley Company.

    Article  CAS  PubMed  Google Scholar 

  18. Li Y, Tang Z, Ye S, Liu Y, Chen J, Xue Q, et al. Establishment of human hepatocellular carcinoma cell line with spontaneous pulmonary metastasis through in vivo selection. Zhonghua Yi Xue Za Zhi. 2002;82:601–5.

    PubMed  Google Scholar 

  19. Li Y, Tang Y, Ye L, Liu B, Liu K, Chen J, et al. Establishment of a hepatocellular carcinoma cell line with unique metastatic characteristics through in vivo selection and screening for metastasis-related genes through cDNA microarray. J Cancer Res Clin Oncol. 2003;129:43–51.

    Article  CAS  PubMed  Google Scholar 

  20. Tao Z-H, Wan J-L, Zeng L-Y, Xie L, Sun H-C, Qin L-X, et al. miR-612 suppresses the invasive-metastatic cascade in hepatocellular carcinoma. J Exp Med. 2013;210:789–803. Rockefeller Univ Press.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Xiao C-L, Tao Z-H, Guo L, Li W-W, Wan J-L, Sun H-C, et al. Isomalto oligosaccharide sulfate inhibits tumor growth and metastasis of hepatocellular carcinoma in nude mice. BMC Cancer. 2011;11:150. BioMed Central Ltd.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Jankowski A, Grinstein S. A noninvasive fluorimetric procedure for measurement of membrane potential. Quantification of the NADPH oxidase-induced depolarization in activated neutrophils. J Biol Chem. 1999;274:26098–104.

    Article  CAS  PubMed  Google Scholar 

  23. Yang Y, Parsons KK, Chi L, Malakauskas SM, Le TH. Glutathione S-transferase-micro1 regulates vascular smooth muscle cell proliferation, migration, and oxidative stress. Hypertension. 2009;54:1360–8. Lippincott Williams & Wilkins.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Zhu X-D, Zhang J-B, Zhuang P-Y, Zhu H-G, Zhang W, Xiong Y-Q, et al. High expression of macrophage colony-stimulating factor in peritumoral liver tissue is associated with poor survival after curative resection of hepatocellular carcinoma. J Clin Oncol. 2008;26:2707–16. American Society of Clinical Oncology.

    Article  PubMed  Google Scholar 

  25. Gill JL, Bishop SC, McCorquodale C, Williams JL, Wiener P. Identification of polymorphisms in the malic enzyme 1, NADP(+)-dependent, cytosolic and nuclear receptor subfamily 0, group B, member 2 genes and their associations with meat and carcass quality traits in commercial Angus cattle. Anim Genet. 2012;43:88–92. Blackwell Publishing Ltd.

    Article  CAS  PubMed  Google Scholar 

  26. Rouaud F, Romero-Perez M, Wang H, Lobysheva I, Ramassamy B, Henry E, et al. Regulation of NADPH-dependent nitric oxide and reactive oxygen species signalling in endothelial and melanoma cells by a photoactive NADPH analogue. Oncotarget. 2014;5:10650–64.

    Article  PubMed  PubMed Central  Google Scholar 

  27. Goodridge AG, Adelman TG. Regulation of malic enzyme synthesis by insulin triiodothyronine, and glucagon in liver cells in culture. J Biol Chem. 1976;251:3027–32.

    CAS  PubMed  Google Scholar 

  28. Istvan ES. Structural mechanism for statin inhibition of 3-hydroxy-3-methylglutaryl coenzyme A reductase. Am Heart J. 2002;144:S27–32.

    Article  CAS  PubMed  Google Scholar 

  29. Wang M, Sun S, Neufeld CI, Perez-Ramirez B, Xu Q. Reactive oxygen species-responsive protein modification and its intracellular delivery for targeted cancer therapy. Angew Chem Int Ed Engl. 2014;53:13444–8. WILEY‐VCH Verlag.

    Article  CAS  PubMed  Google Scholar 

  30. Liu-Smith F, Dellinger R, Meyskens FL. Updates of reactive oxygen species in melanoma etiology and progression. Arch Biochem Biophys. 2014;563C:51–5.

    Article  Google Scholar 

  31. Wei S, Wang Y, Chai Q, Fang Q, Zhang Y, Wang J. Potential crosstalk of Ca2+−ROS-dependent mechanism involved in apoptosis of Kasumi-1 cells mediated by heme oxygenase-1 small interfering RNA. Int J Oncol. 2014;45:2373–84. Spandidos Publications.

    CAS  PubMed  Google Scholar 

  32. Esmaeili MA, Farimani MM, Kiaei M. Anticancer effect of calycopterin via PI3K/Akt and MAPK signaling pathways, ROS-mediated pathway and mitochondrial dysfunction in hepatoblastoma cancer (HepG2) cells. Mol Cell Biochem. 2014;397:17–31. Springer US.

    Article  CAS  PubMed  Google Scholar 

  33. Yang M-H, Chen C-L, Chau G-Y, Chiou S-H, Su C-W, Chou T-Y, et al. Comprehensive analysis of the independent effect of twist and snail in promoting metastasis of hepatocellular carcinoma. Hepatology. 2009;50:1464–74. Wiley Subscription Services, Inc., A Wiley Company.

    Article  CAS  PubMed  Google Scholar 

  34. Li J, Dong L, Wei D, Wang X, Zhang S, Li H. Fatty acid synthase mediates the epithelial-mesenchymal transition of breast cancer cells. Int J Biol Sci. 2014;10:171–80.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Liu Y, Liu Y, Yan X, Xu Y, Luo F, Ye J, et al. HIFs enhance the migratory and neoplastic capacities of hepatocellular carcinoma cells by promoting EMT. Tumour Biol. 2014;35:8103–14. Springer Netherlands.

    Article  CAS  PubMed  Google Scholar 

  36. Das TP, Suman S, Damodaran C. Induction of reactive oxygen species generation inhibits epithelial–mesenchymal transition and promotes growth arrest in prostate cancer cells. Mol Carcinog. 2014;53:537–47.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The project was jointly supported by the National Science Foundation of China (81272437, 81472675). We thank Mengxin Tian for providing clinical data of HCC patients.

Conflicts of interest

None

Authors’ Contributions

D Wen, DL Liu, and WZ Wu conceived and designed the study. D Wen, DL Liu, J Tang, LL Dong, Y Liu, JL Wan, and ZH Tang performed the experiments including IHC, RT-PCR, Western blotting, and in vivo assays. D Wen, and DL Liu performed NADPH quantification assays. D Wen, DL Liu, and WZ Wu analyzed the data and prepared the manuscript. L Wang, HC Sun, and J Fan participated in the study design. All authors read and approved the final manuscript.

Author information

Author notes

    Authors and Affiliations

    1. Liver Cancer Institute, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai, 2000032, China

      Duo Wen, Dongli Liu, Jun Tang, Lili Dong, Yang Liu, Zhonghua Tao, Jinliang Wan, Dongmei Gao, Lu Wang, Huichuan Sun, Jia Fan & Weizhong Wu

    2. Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Shanghai, 200032, China

      Duo Wen, Dongli Liu, Jun Tang, Lili Dong, Yang Liu, Zhonghua Tao, Jinliang Wan, Dongmei Gao, Lu Wang, Huichuan Sun, Jia Fan & Weizhong Wu

    3. Institute of Biomedical Sciences, Fudan University, Shanghai, 200032, China

      Jia Fan

    Authors
    1. Duo Wen
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Dongli Liu
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Jun Tang
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Lili Dong
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. Yang Liu
      View author publications

      You can also search for this author in PubMed Google Scholar

    6. Zhonghua Tao
      View author publications

      You can also search for this author in PubMed Google Scholar

    7. Jinliang Wan
      View author publications

      You can also search for this author in PubMed Google Scholar

    8. Dongmei Gao
      View author publications

      You can also search for this author in PubMed Google Scholar

    9. Lu Wang
      View author publications

      You can also search for this author in PubMed Google Scholar

    10. Huichuan Sun
      View author publications

      You can also search for this author in PubMed Google Scholar

    11. Jia Fan
      View author publications

      You can also search for this author in PubMed Google Scholar

    12. Weizhong Wu
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to Weizhong Wu.

    Additional information

    Duo Wen and Dongli Liu contributed equally to this work.

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Wen, D., Liu, D., Tang, J. et al. Malic enzyme 1 induces epithelial–mesenchymal transition and indicates poor prognosis in hepatocellular carcinoma. Tumor Biol. 36, 6211–6221 (2015). https://doi.org/10.1007/s13277-015-3306-5

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s13277-015-3306-5

    Keywords

    Search

    Navigation