Sublethal heat treatment promotes epithelial-mesenchymal transition and enhances the malignant potential of hepatocellular carcinoma

Authors

  • Shuhei Yoshida,

    1. Division of Gastroenterology and Liver Center, Beth Israel Deaconess Medical Center, Boston, MA
    Search for more papers by this author
  • Miroslaw Kornek,

    1. Division of Gastroenterology and Liver Center, Beth Israel Deaconess Medical Center, Boston, MA
    2. Department of Medicine II, Saarland University Medical Center, Homburg, Germany
    Search for more papers by this author
  • Naoki Ikenaga,

    1. Division of Gastroenterology and Liver Center, Beth Israel Deaconess Medical Center, Boston, MA
    Search for more papers by this author
  • Moritz Schmelzle,

    1. Transplantation Institute, Department of Medicine and Surgery, Beth Israel Deaconess Medical Center, Boston, MA
    2. Department of Surgery and Translational Centre for Regenerative Medicine, University of Leipzig, Germany
    Search for more papers by this author
  • Ryota Masuzaki,

    1. Division of Gastroenterology and Liver Center, Beth Israel Deaconess Medical Center, Boston, MA
    2. Transplantation Institute, Department of Medicine and Surgery, Beth Israel Deaconess Medical Center, Boston, MA
    Search for more papers by this author
  • Eva Csizmadia,

    1. Transplantation Institute, Department of Medicine and Surgery, Beth Israel Deaconess Medical Center, Boston, MA
    Search for more papers by this author
  • Yan Wu,

    1. Transplantation Institute, Department of Medicine and Surgery, Beth Israel Deaconess Medical Center, Boston, MA
    Search for more papers by this author
  • Simon C. Robson,

    1. Division of Gastroenterology and Liver Center, Beth Israel Deaconess Medical Center, Boston, MA
    2. Transplantation Institute, Department of Medicine and Surgery, Beth Israel Deaconess Medical Center, Boston, MA
    Search for more papers by this author
  • Detlef Schuppan

    Corresponding author
    1. Division of Gastroenterology and Liver Center, Beth Israel Deaconess Medical Center, Boston, MA
    2. Molecular and Translational Medicine, Dept. Medicine I, University of Mainz Medical School, Mainz, Germany
    • Address reprint requests to: Detlef Schuppan, M.D., Ph.D., Division of Gastroenterology and Liver Center, Beth Israel Deaconess Medical Center, Harvard Medical School, 330 Brookline Avenue, Boston, MA 02115. E-mail: dschuppan@bidmc.harvard.edu; fax: 617-667-2767.

    Search for more papers by this author

  • Potential conflict of interest: Nothing to report.

  • This work was supported by the National Institute of Diabetes and Digestive and Kidney Diseases (grant nos.: 1 R21 DK075857-01A2 and U19 AI066313-05; to D.S.) and the German Research Foundation (DFG; grant no.: KO4103/1-1; to M.K.). M.S. also acknowledges grant support from the American Liver Foundation, the DFG (SCHM 2661/1-1 and 2661/1-2), and the German Federal Ministry of Education and Research (BMBF; PtJ-Bio, 0315883).

  • Some aspects of this study were presented at an oral sessions of Digestive Disease Week 2011 (Abstract no. 896) and at poster sessions of the American Association for the Study of Liver Diseases 2012 (Abstract no. 887) and 2013 (Abstract no. 1804).

Abstract

Radiofrequency ablation (RFA) is a potentially curative therapy for hepatocellular carcinoma (HCC). However, incomplete RFA can induce accelerated invasive growth at the periphery. The mechanisms underlying the RFA-induced tumor promotion remain largely unexplored. Three human HCC cell lines were exposed to 45°C-55°C for 10 minutes, simulating the marginal zone of RFA treatment. At 5-12 days post-treatment cell proliferation, parameters of epithelial-mesenchymal transition (EMT), and activation of mitogen-activated protein kinases were analyzed. Livers from patients with viral hepatitis without and with HCC (n = 114) were examined to confirm the relevance of altered kinase patterns. In vivo tumorigenic potential of heat-treated versus untreated HCC cells was studied in nude mice. Heating to 55°C killed all HCC cells, whereas 65%-85% of cells survived 48°C-50°C, developing spindle-like morphology and expressing CD133, cytokeratin (CK)7, CK19, procollagen-α1(I), and Snail at day 5 after heat exposure, which returned to baseline at day 12. Heat-exposed HCC cells showed enhanced proliferation and prominent activation of p46-Shc (Src homology and collagen) and downstream extracellular signal-related kinase (Erk)1/2. In patients, Shc expression correlated with malignant potential and overall survival. Blocking Erk1/2 reduced proliferation and EMT-like changes of heat-treated HCC cells. Implantation of heat-exposed HEPG2 cells into nude mice induced significantly larger, more aggressive tumors than untreated cells. Conclusions: Sublethal heat treatment skews HCC cells toward EMT and transforms them to a progenitor-like, highly proliferative cellular phenotype in vitro and in vivo, which is driven significantly by p46Shc-Erk1/2. Suboptimal RFA accelerates HCC growth and spread by transiently inducing an EMT-like, more aggressive cellular phenotype. (Hepatology 2013;58:1667–1680)

Ancillary