Potential conflict of interest: Nothing to report.
Sublethal heat treatment promotes epithelial-mesenchymal transition and enhances the malignant potential of hepatocellular carcinoma
Article first published online: 19 SEP 2013
© 2013 by the American Association for the Study of Liver Diseases
Volume 58, Issue 5, pages 1667–1680, November 2013
How to Cite
Yoshida, S., Kornek, M., Ikenaga, N., Schmelzle, M., Masuzaki, R., Csizmadia, E., Wu, Y., Robson, S. C. and Schuppan, D. (2013), Sublethal heat treatment promotes epithelial-mesenchymal transition and enhances the malignant potential of hepatocellular carcinoma. Hepatology, 58: 1667–1680. doi: 10.1002/hep.26526
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).
- Issue published online: 30 OCT 2013
- Article first published online: 19 SEP 2013
- Accepted manuscript online: 31 MAY 2013 08:42AM EST
- Manuscript Accepted: 10 MAY 2013
- Manuscript Received: 21 NOV 2012
- National Institute of Diabetes and Digestive and Kidney Diseases. Grant Numbers: 1 R21 DK075857-01A2, U19 AI066313-05
- German Research Foundation (DFG). Grant Number: KO4103/1-1
- American Liver Foundation, the DFG. Grant Numbers: SCHM 2661/1-1, 2661/1-2
- German Federal Ministry of Education and Research (BMBF; PtJ-Bio). Grant Number: 0315883
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)