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Experimental Therapeutics, Molecular Targets, and Chemical Biology |
1 Department of Radiation Genetics; 2 Department of Neurosurgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan; 3 Department of Radiation and Cellular Oncology, University of Chicago, Chicago, Illinois; 4 Immunology and Molecular Genetics, Kawasaki Medical School, Okayama, Japan; and 5 Forschungszentrum für Umwelt und Gesundheit, Institute for Molecular Radiobiology, Neuherberg-Munich, Germany
Requests for reprints: Eiichiro Sonoda, Department of Radiation Genetics, Graduate School of Medicine, Kyoto University, Konoe Yoshida, Sakyo-ku, Kyoto 606-8501, Japan. Phone: 81-75-753-4412; Fax: 81-75-753-4419; E-mail: esonoda{at}rg.med.kyoto-u.ac.jp.
Cross-linking agents that induce DNA interstrand cross-links (ICL) are widely used in anticancer chemotherapy. Yeast genetic studies show that nucleotide excision repair (NER), Rad6/Rad18-dependent postreplication repair, homologous recombination, and cell cycle checkpoint pathway are involved in ICL repair. To study the contribution of DNA damage response pathways in tolerance to cross-linking agents in vertebrates, we made a panel of gene-disrupted clones from chicken DT40 cells, each defective in a particular DNA repair or checkpoint pathway, and measured the sensitivities to cross-linking agents, including cis-diamminedichloroplatinum (II) (cisplatin), mitomycin C, and melphalan. We found that cells harboring defects in translesion DNA synthesis (TLS), Fanconi anemia complementation groups (FANC), or homologous recombination displayed marked hypersensitivity to all the cross-linking agents, whereas NER seemed to play only a minor role. This effect of replication-dependent repair pathways is distinctively different from the situation in yeast, where NER seems to play a major role in dealing with ICL. Cells deficient in Rev3, the catalytic subunit of TLS polymerase Pol
, showed the highest sensitivity to cisplatin followed by fanc-c. Furthermore, epistasis analysis revealed that these two mutants work in the same pathway. Our genetic comprehensive study reveals a critical role for DNA repair pathways that release DNA replication block at ICLs in cellular tolerance to cross-linking agents and could be directly exploited in designing an effective chemotherapy. (Cancer Res 2005; 65(24): 11704-11)
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