title: Epithelial to mesenchymal transition in the liver field: the double face of Everolimus in vitro.
abstract: BACKGROUND
  Everolimus (EVE), a mammalian target of rapamycin inhibitor, has been proposed as liver transplant immunosuppressive drug, gaining wide interest also for the treatment of cancer. Although an appropriate tolerance, it may induce several adverse effects, such as fibro-interstitial pneumonitis due to the acquisition of activated myofibroblasts. The exact molecular mechanism associated with epithelial to mesenchymal transition (EMT) may be crucial also in the liver context. This work examines the role and the molecular mediators of EMT in hepatic stellate cell (HSC) and human liver cancer cells (HepG2) and the potential role of EVE to maintain the epithelial phenotype rather than to act as a potential initiators of EMT.
  
  
  METHODS
  Real time-PCR and western blot have been used to assess the capability of EVE at low-therapeutic (10 nM) and high (100 nM) dose to induce an in vitro EMT in HSC and HepG2.
  
  
  RESULTS
  Biomolecular experiments demonstrated that low concentration of EVE (10 nM) did not modify the gene expression of alpha-smooth muscle actin (α-SMA), Vimentin (VIM), Fibronectin (FN) in both HSC and HepG2 cells, whereas EVE at 100 nM induced a significant over-expression of all the three above-mentioned genes and an increment of α-SMA and FN protein levels. Additionally, 100 nM of EVE induced a significant phosphorylation of AKT and an up-regulation of TGF-β expression in HSC and HepG2 cells.
  
  
  CONCLUSIONS
  Our data, although obtained in an in vitro model, revealed, for the first time, that high concentration of EVE may induce EMT in liver cells confirming previous published evidences obtained in renal cells. Additionally, they suggested that mTOR-I should be administered at the lowest dose able to maximize their important and specific therapeutic properties minimizing or avoiding fibrosis-related adverse effects.
  
  
  CONCLUSIONS
  In summary, if confirmed by additional studies, our results could be useful for researchers to standardize new therapeutic immunosuppressive and anticancer drugs protocols.
authors.affiliation: Deparment of Medicine, Renal Unit, University Hospital of Verona, 37126, Verona, Italy. valentina.masola@unipd.it.
authors.forename: Valentina
authors.initials: V
authors.lastname: Masola
authors.affiliation: Department of General Surgery and Odontoiatrics, Liver Transplant Unit, University Hospital of Verona, 37126, Verona, Italy. amedeo.carraro@ospedaleuniverona.it.
authors.forename: Amedeo
authors.initials: A
authors.lastname: Carraro
authors.affiliation: Deparment of Medicine, Renal Unit, University Hospital of Verona, 37126, Verona, Italy. gianluigi.zaza@univr.it.
authors.forename: Gianluigi
authors.initials: G
authors.lastname: Zaza
authors.affiliation: Deparment of Medicine, Renal Unit, University Hospital of Verona, 37126, Verona, Italy. gloria.bellin@unipd.it.
authors.forename: Gloria
authors.initials: G
authors.lastname: Bellin
authors.affiliation: Department of General Surgery and Odontoiatrics, Liver Transplant Unit, University Hospital of Verona, 37126, Verona, Italy. umbertomontin@ospedaleuniverona.it.
authors.forename: Umberto
authors.initials: U
authors.lastname: Montin
authors.affiliation: Department of General Surgery and Odontoiatrics, Liver Transplant Unit, University Hospital of Verona, 37126, Verona, Italy. paolavioli@ospedaleuniverona.it.
authors.forename: Paola
authors.initials: P
authors.lastname: Violi
authors.affiliation: Deparment of Medicine, Renal Unit, University Hospital of Verona, 37126, Verona, Italy. antonio.lupo@univr.it.
authors.forename: Antonio
authors.initials: A
authors.lastname: Lupo
authors.affiliation: Department of General Surgery and Odontoiatrics, Liver Transplant Unit, University Hospital of Verona, 37126, Verona, Italy. umberto.tedeschi@ospedaeuniverona.it1.
authors.forename: Umberto
authors.initials: U
authors.lastname: Tedeschi
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects*; D000068338:Everolimus / Q000008:administration & dosage / Q000494:pharmacology*; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects*; D056945:Hep G2 Cells; D055166:Hepatic Stellate Cells / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D007166:Immunosuppressive Agents / Q000008:administration & dosage / Q000494:pharmacology*; D010641:Phenotype; D010766:Phosphorylation / Q000187:drug effects; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D016212:Transforming Growth Factor beta / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects; D014746:Vimentin / Q000235:genetics
chemical_list: D000199:Actins; D000068338:Everolimus; D005353:Fibronectins; D007166:Immunosuppressive Agents; D051057:Proto-Oncogene Proteins c-akt; D016212:Transforming Growth Factor beta; D014746:Vimentin; C541116:ACTA2 protein, human; C498862:Acta2 protein, rat