Simvastatin and metformin inhibit cell growth in hepatitis C virus infected cells via mTOR increasing PTEN and autophagy

PLoS One. 2018 Jan 31;13(1):e0191805. doi: 10.1371/journal.pone.0191805. eCollection 2018.

Abstract

Hepatitis C virus (HCV) infection has been related to increased risk of development of hepatocellular carcinoma (HCC) while metformin (M) and statins treatment seemed to protect against HCC development. In this work, we aim to identify the mechanisms by which metformin and simvastatin (S) could protect from liver cancer. Huh7.5 cells were infected with HCV particles and treated with M+S. Human primary hepatocytes were treated with M+S. Treatment with both drugs inhibited Huh7.5 cell growth and HCV infection. In non-infected cells S increased translational controlled tumor protein (TCTP) and phosphatase and tensin homolog (PTEN) proteins while M inhibited mammalian target of rapamycin (mTOR) and TCTP. Simvastatin and metformin co-administered down-regulated mTOR and TCTP, while PTEN was increased. In cells infected by HCV, mTOR, TCTP, p62 and light chain 3B II (LC3BII) were increased and PTEN was decreased. S+M treatment increased PTEN, p62 and LC3BII in Huh7.5 cells. In human primary hepatocytes, metformin treatment inhibited mTOR and PTEN, but up-regulated p62, LC3BII and Caspase 3. In conclusion, simvastatin and metformin inhibited cell growth and HCV infection in vitro. In human hepatocytes, metformin increased cell-death markers. These findings suggest that M+S treatment could be useful in therapeutic prevention of HCV-related hepatocellular carcinoma.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autophagy / drug effects
  • Biomarkers, Tumor / genetics
  • Biomarkers, Tumor / metabolism
  • Carcinoma, Hepatocellular / prevention & control
  • Caspase 3 / metabolism
  • Cell Line
  • Cell Proliferation / drug effects
  • Cells, Cultured
  • Drug Therapy, Combination
  • Gene Expression / drug effects
  • Hepacivirus / drug effects*
  • Hepacivirus / pathogenicity
  • Hepatitis C, Chronic / complications
  • Hepatitis C, Chronic / drug therapy
  • Hepatocytes / drug effects*
  • Hepatocytes / metabolism
  • Hepatocytes / virology*
  • Humans
  • Liver Neoplasms / prevention & control
  • Metformin / administration & dosage*
  • Microtubule-Associated Proteins / metabolism
  • PTEN Phosphohydrolase / metabolism*
  • Simvastatin / administration & dosage*
  • TOR Serine-Threonine Kinases / genetics*
  • TOR Serine-Threonine Kinases / metabolism
  • Tumor Protein, Translationally-Controlled 1

Substances

  • Biomarkers, Tumor
  • MAP1LC3B protein, human
  • Microtubule-Associated Proteins
  • TPT1 protein, human
  • Tumor Protein, Translationally-Controlled 1
  • Metformin
  • Simvastatin
  • MTOR protein, human
  • TOR Serine-Threonine Kinases
  • PTEN Phosphohydrolase
  • PTEN protein, human
  • CASP3 protein, human
  • Caspase 3

Grant support

This work was supported by the Spanish Ministry of Economy, Innovation and Competitivity, Instituto de Salud Carlos III, grant numbers PI13/01192, PI14/01349, co-funded by European Union (ERDF/ESF, “Investing in your future”) and Consejería de Salud de la Junta de Andalucía (PI-0892-2012). JA Del Campo was supported by Nicolás Monardes Program from Servicio Andaluz de Salud (SAS).