Targeting glucosylceramide synthase upregulation reverts sorafenib resistance in experimental hepatocellular carcinoma

Oncotarget. 2016 Feb 16;7(7):8253-67. doi: 10.18632/oncotarget.6982.

Abstract

Evasive mechanisms triggered by the tyrosine kinase inhibitor sorafenib reduce its efficacy in hepatocellular carcinoma (HCC) treatment. Drug-resistant cancer cells frequently exhibit sphingolipid dysregulation, reducing chemotherapeutic cytotoxicity via the induction of ceramide-degrading enzymes. However, the role of ceramide in sorafenib therapy and resistance in HCC has not been clearly established. Our data reveals that ceramide-modifying enzymes, particularly glucosylceramide synthase (GCS), are upregulated during sorafenib treatment in hepatoma cells (HepG2 and Hep3B), and more importantly, in sorafenib-resistant cell lines. GCS silencing or pharmacological GCS inhibition sensitized hepatoma cells to sorafenib exposure. GCS inhibition, combined with sorafenib, triggered cytochrome c release and ATP depletion in sorafenib-treated hepatoma cells, leading to mitochondrial cell death after energetic collapse. Conversely, genetic GCS overexpression increased sorafenib resistance. Of interest, GCS inhibition improved sorafenib effectiveness in a xenograft mouse model, recovering drug sensitivity of sorafenib-resistant tumors in mice. In conclusion, our results reveal GCS induction as a mechanism of sorafenib resistance, suggesting that GCS targeting may be a novel strategy to increase sorafenib efficacy in HCC management, and point to target the mitochondria as the subcellular location where sorafenib therapy could be potentiated.

Keywords: ceramide; chemotherapy; liver cancer; mitochondria; mouse model.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Blotting, Western
  • Carcinoma, Hepatocellular / drug therapy*
  • Carcinoma, Hepatocellular / enzymology
  • Carcinoma, Hepatocellular / pathology
  • Cell Proliferation / drug effects
  • Ceramides / metabolism
  • Drug Resistance, Neoplasm / drug effects*
  • Fatty Acids, Monounsaturated / pharmacology
  • Gene Expression Regulation, Enzymologic / drug effects*
  • Gene Expression Regulation, Neoplastic / drug effects
  • Glucosyltransferases / antagonists & inhibitors*
  • Glucosyltransferases / genetics
  • Humans
  • Immunoenzyme Techniques
  • Immunosuppressive Agents / pharmacology
  • Liver Neoplasms, Experimental / drug therapy*
  • Liver Neoplasms, Experimental / enzymology
  • Liver Neoplasms, Experimental / pathology
  • Male
  • Mice
  • Mice, Nude
  • Niacinamide / analogs & derivatives*
  • Niacinamide / pharmacology
  • Phenylurea Compounds / pharmacology*
  • Protein Kinase Inhibitors / pharmacology
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction / drug effects
  • Sorafenib
  • Tumor Cells, Cultured
  • Xenograft Model Antitumor Assays

Substances

  • Ceramides
  • Fatty Acids, Monounsaturated
  • Immunosuppressive Agents
  • Phenylurea Compounds
  • Protein Kinase Inhibitors
  • RNA, Messenger
  • Niacinamide
  • Sorafenib
  • Glucosyltransferases
  • ceramide glucosyltransferase
  • thermozymocidin