title: Quantitative proteomic analysis of anticancer drug RH1 resistance in liver carcinoma.
abstract: Acquired resistance of tumor cells to the therapeutic treatment is a major challenge in virtually any chemotherapy. A novel anticancer agent 2,5-diaziridinyl-3-(hydroxymethyl)-6-methyl-1,4-benzoquinone (RH1) is designed to be activated by
  quinone oxidoreductase, an enzyme expressed at high levels in many types of tumors. Here we investigated the potential mechanisms of acquired RH1 drug resistance in cancer cells by applying high-throughput differential quantitative proteomic analysis of the newly established RH1-resistant hepatoma cell lines. Over 400 proteins display significantly altered levels between drug-sensitive and drug-resistant cell lines. Differentially expressed proteins were clustered into more than 14 groups according to their functional annotation and protein-protein interactions. Bioinformatic analysis highlights the biological processes that might be responsible for acquired resistance to RH1. The level of several xenobiotic metabolism enzymes (total n=17) involved in RH1 activation and detoxification is decreased (Nqo1, catalase, Gst, Gsr), corresponding with the decrease in their catalytic activity. The altered biological processes also include the decrease of cell cycle positive regulators (n=15) and the increase of DNA repair proteins (n=5) as well as annexin family members (n=5) in the RH1-resistant cells. Drug-resistant hepatoma cell proteomes are also distinguished by the altered level of proteins involved in energy production and metabolism (n=55). Our data provide the basis for in-depth study of molecular mechanisms of tumor cell resistance to the promising anticancer drug RH1 enabling the further validation of protein biomarkers for the drug insusceptibility and of potential secondary pharmacological targets of RH1 resistant cells.
authors.affiliation: Vilnius University Institute of Biochemistry, Vilnius, Lithuania. Electronic address: marija.ger@bchi.vu.lt.
authors.forename: Marija
authors.initials: M
authors.lastname: Ger
authors.affiliation: Vilnius University Institute of Biochemistry, Vilnius, Lithuania. Electronic address: algirdas.kaupinis@gf.vu.lt.
authors.forename: Algirdas
authors.initials: A
authors.lastname: Kaupinis
authors.affiliation: State Research Institute Centre for Innovative Medicine, Vilnius, Lithuania. Electronic address: a.ceniene@imcentras.lt.
authors.forename: Ausra
authors.initials: A
authors.lastname: Nemeikaite-Ceniene
authors.affiliation: Vilnius University Institute of Biochemistry, Vilnius, Lithuania. Electronic address: jonas.sarlauskas@bchi.vu.lt.
authors.forename: Jonas
authors.initials: J
authors.lastname: Sarlauskas
authors.affiliation: Vilnius University Institute of Biochemistry, Vilnius, Lithuania; Swiss Institute of Bioinformatics, Geneva, Switzerland; MAP Kinase Resource, Bern, Switzerland. Electronic address: j.cicenas@mapkinases.eu.
authors.forename: Jonas
authors.initials: J
authors.lastname: Cicenas
authors.affiliation: Vilnius University Institute of Biochemistry, Vilnius, Lithuania. Electronic address: narimantas.cenas@bchi.vu.lt.
authors.forename: Narimantas
authors.initials: N
authors.lastname: Cenas
authors.affiliation: Vilnius University Institute of Biochemistry, Vilnius, Lithuania. Electronic address: mindaugas.valius@bchi.vu.lt.
authors.forename: Mindaugas
authors.initials: M
authors.lastname: Valius
mesh_terms: D001388:Aziridines / Q000008:administration & dosage*; D016227:Benzoquinones / Q000008:administration & dosage*; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy* / Q000235:genetics; D045744:Cell Line, Tumor; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D008113:Liver Neoplasms / Q000188:drug therapy* / Q000235:genetics; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000235:genetics; D020543:Proteome / Q000187:drug effects / Q000235:genetics; D040901:Proteomics
chemical_list: C117776:2,5-diaziridinyl-3-(hydroxymethyl)-6-methyl-1,4-benzoquinone; D001388:Aziridines; D016227:Benzoquinones; D009363:Neoplasm Proteins; D020543:Proteome
keywords: Cancer drug resistance; Chemotherapy; High-throughput proteomics; NQO1; RH1