title: Influence of fatty acid synthase inhibitor orlistat on the DNA repair enzyme O6-methylguanine-DNA methyltransferase in human normal or malignant cells in vitro.
abstract: Tetrahydrolipstatin (orlistat), an inhibitor of lipases and fatty acid synthase, is used orally for long-term treatment of obesity. Although the drug possesses striking antitumor activities in vitro against human cancer cells and in vitro and in vivo against animal tumors, it also induces precancerous lesions in rat colon. Therefore, we tested the in vitro effect of orlistat on the expression of O6-methylguanine-DNA methyltransferase (MGMT), a DNA repair enzyme that plays an essential role in the control of mutagenesis and carcinogenesis. Western blot analysis demonstrated that 2-day continuous exposure to 40 µM orlistat did not affect MGMT levels in a human melanoma cell line, but downregulated the repair protein by 30-70% in human peripheral blood mononuclear cells, in two leukemia and two colon cancer cell lines. On the other hand, orlistat did not alter noticeably MGMT mRNA expression. Differently from lomeguatrib (a false substrate, strong inhibitor of MGMT) orlistat did not reduce substantially MGMT function after 2-h exposure of target cells to the agent, suggesting that this drug is not a competitive inhibitor of the repair protein. Combined treatment with orlistat and lomeguatrib showed additive reduction of MGMT levels. More importantly, orlistat-mediated downregulation of MGMT protein expression was markedly amplified when the drug was combined with a DNA methylating agent endowed with carcinogenic properties such as temozolomide. In conclusion, even if orlistat is scarcely absorbed by oral route, it is possible that this drug could reduce local MGMT-mediated protection against DNA damage provoked by DNA methylating compounds on gastrointestinal tract epithelial cells, thus favoring chemical carcinogenesis.
authors.affiliation: Department of Systems Medicine, University of Rome 'Tor Vergata', I-00133 Rome, Italy.
authors.forename: Giorgia
authors.initials: G
authors.lastname: Cioccoloni
authors.affiliation: Laboratory of Molecular Oncology, Istituto Dermopatico dell'Immacolata-IRCCS, I-00167 Rome, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Bonmassar
authors.affiliation: Laboratory of Molecular Oncology, Istituto Dermopatico dell'Immacolata-IRCCS, I-00167 Rome, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Pagani
authors.affiliation: Laboratory of Molecular Oncology, Istituto Dermopatico dell'Immacolata-IRCCS, I-00167 Rome, Italy.
authors.forename: Simona
authors.initials: S
authors.lastname: Caporali
authors.affiliation: Institute of Translational Pharmacology (IFT), National Research Council (CNR), I-00133 Rome, Italy.
authors.forename: Maria Pia
authors.initials: MP
authors.lastname: Fuggetta
authors.affiliation: Institute of Translational Pharmacology (IFT), National Research Council (CNR), I-00133 Rome, Italy.
authors.forename: Enzo
authors.initials: E
authors.lastname: Bonmassar
authors.affiliation: Laboratory of Molecular Oncology, Istituto Dermopatico dell'Immacolata-IRCCS, I-00167 Rome, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: D'Atri
authors.affiliation: Department of Systems Medicine, University of Rome 'Tor Vergata', I-00133 Rome, Italy.
authors.forename: Angelo
authors.initials: A
authors.lastname: Aquino
mesh_terms: D045744:Cell Line, Tumor; D015254:DNA Modification Methylases / Q000235:genetics / Q000378:metabolism*; D045643:DNA Repair Enzymes / Q000235:genetics / Q000378:metabolism*; D003606:Dacarbazine / Q000031:analogs & derivatives; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D045325:HCT116 Cells; D019073:HT29 Cells; D006801:Humans; D066298:In Vitro Techniques; D007783:Lactones / Q000494:pharmacology*; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000201:enzymology*; D009369:Neoplasms / Q000201:enzymology* / Q000235:genetics; D000077403:Orlistat; D011687:Purines / Q000494:pharmacology; D000077204:Temozolomide; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D004791:Enzyme Inhibitors; D007783:Lactones; D011687:Purines; D025521:Tumor Suppressor Proteins; D003606:Dacarbazine; D000077403:Orlistat; D015254:DNA Modification Methylases; C515491:MGMT protein, human; D045643:DNA Repair Enzymes; C521206:lomeguatrib; D000077204:Temozolomide