title: Expression of domains for protein-protein interaction of nucleotide excision repair proteins modifies cancer cell sensitivity to platinum derivatives and genomic stability.
abstract: Nucleotide excision repair (NER) is involved in the repair of DNA damage caused by platinum derivatives and has been shown to decrease the cytotoxic activity of these drugs. Because protein-protein interactions are essential for NER activity, we transfected human cancer cell lines (A549 and HCT116) with plasmids coding the amino acid sequences corresponding to the interacting domains between excision repair cross-complementation group 1 (ERCC1) and xeroderma pigmentosum, complementation group A (XPA), as well as ERCC1 and xeroderma pigmentosum, complementation group F (XPF), all NER proteins. Using the 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and annexin V staining, we showed that transfected A549 cells were sensitized 1.2-2.2-fold to carboplatin and that transfected HCT116 cells were sensitized 1.4-5.4-fold to oxaliplatin in vitro. In addition, transfected cells exhibited modified in vivo sensitivity to the same drugs. Finally, in particular cell models of the interaction between ERCC1 and XPF, DNA repair was decreased, as evidenced by increased phosphorylation of the histone 2AX after exposure to mitomycin C, and genomic instability was increased, as determined by comparative genomic hybridization studies. The results indicate that the interacting peptides act as dominant negatives and decrease NER activity through inhibition of protein-protein interactions.
authors.affiliation: Université de Lyon, Lyon, France; Université de Lyon 1, Lyon, France; Cancer Research Center in Lyon, INSERM U1052, Lyon, France; Cancer Research Center in Lyon, CNRS UMR 5286, Lyon, France.
authors.forename: Lars Petter
authors.initials: LP
authors.lastname: Jordheim
authors.forename: Emeline
authors.initials: E
authors.lastname: Cros-Perrial
authors.forename: Eva-Laure
authors.initials: EL
authors.lastname: Matera
authors.forename: Karima
authors.initials: K
authors.lastname: Bouledrak
authors.forename: Charles
authors.initials: C
authors.lastname: Dumontet
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D016190:Carboplatin / Q000494:pharmacology; D045744:Cell Line, Tumor; D055028:Comparative Genomic Hybridization / Q000379:methods; D004249:DNA Damage / Q000187:drug effects / Q000235:genetics; D004260:DNA Repair / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics; D004720:Endonucleases / Q000235:genetics; D042822:Genomic Instability / Q000235:genetics*; D045325:HCT116 Cells; D006801:Humans; D009369:Neoplasms / Q000188:drug therapy* / Q000235:genetics; D009944:Organoplatinum Compounds / Q000494:pharmacology*; D000077150:Oxaliplatin; D010455:Peptides / Q000235:genetics; D010766:Phosphorylation / Q000187:drug effects / Q000235:genetics; D054730:Protein Interaction Domains and Motifs / Q000235:genetics*; D011506:Proteins / Q000235:genetics*; D014162:Transfection / Q000379:methods; D014983:Xeroderma Pigmentosum / Q000235:genetics
chemical_list: D000970:Antineoplastic Agents; D004268:DNA-Binding Proteins; D009944:Organoplatinum Compounds; D010455:Peptides; D011506:Proteins; C095888:xeroderma pigmentosum group F protein; D000077150:Oxaliplatin; D016190:Carboplatin; C071447:ERCC1 protein, human; D004720:Endonucleases
keywords: cisplatin; complementation group A (XPA); complementation group F (XPF); excision repair cross-complementation group 1 (ERCC1); resistance; sensitization; xeroderma pigmentosum