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Expression of domains for protein–protein interaction of nucleotide excision repair proteins modifies cancer cell sensitivity to platinum derivatives and genomic stability
Author(s) -
Jordheim Lars Petter,
CrosPerrial Emeline,
Matera EvaLaure,
Bouledrak Karima,
Dumontet Charles
Publication year - 2014
Publication title -
clinical and experimental pharmacology and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.752
H-Index - 103
eISSN - 1440-1681
pISSN - 0305-1870
DOI - 10.1111/1440-1681.12282
Subject(s) - nucleotide , nucleotide excision repair , platinum , cancer cell , protein expression , cancer , chemistry , biology , cancer research , gene , genetics , dna repair , biochemistry , catalysis
Summary 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 HCT 116) with plasmids coding the amino acid sequences corresponding to the interacting domains between excision repair cross‐complementation group 1 ( ERCC 1) and xeroderma pigmentosum, complementation group A ( XPA ), as well as ERCC 1 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 HCT 116 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 ERCC 1 and XPF , DNA repair was decreased, as evidenced by increased phosphorylation of the histone 2 AX 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.