Deficient gene specific repair of cisplatin-induced lesions in Xeroderma pigmentosum and Fanconi's anemia cell lines.
Cisplatin is a chemotherapeutic agent known to cause DNA damage. The cytotoxicity of this drug is believed to result from the formation of DNA intrastrand adducts (IA) and DNA interstrand crosslinks (ICL). While there are many studies on DNA repair of cisplatin damage at the overall level of the genome in various human cell lines, there is little information on the gene-specific repair. In this report, we have measured the formation and repair of cisplatin induced DNA adducts in the dihydrofolate reductase (DHFR) and ribosomal RNA (rRNA) genes in three cell lines: normal human fibroblasts, Fanconi's anemia complementation group A (FAA) and Xeroderma pigmentosum complementation group A (XPA). It is generally thought that XPA cells lack nucleotide excision repair and that FAA cells are deficient in the repair of DNA ICL. We find that normal human fibroblast cells repair 84% of the ICL in the DHFR gene after 24 h, whereas XPA and FAA cell lines only repaired 32 and 50% of the ICL respectively. Furthermore, 69% of the cisplatin IA in the DHFR gene were repaired in 24 h in normal human fibroblasts compared to 22% for XPA and 24% for FAA cells. The repair of the rRNA gene was less efficient than in the DHFR gene, but the relative pattern between the different cell lines was similar to that of the DHFR gene. We thus find that FAA cells are deficient not only in the gene specific repair of cisplatin ICL, but also in the gene specific repair of the more common cisplatin IA. XPA cells are normally thought to be without any nucleotide excision repair capacity, but our data could support a slight ICL unhooking activity.
Laboratory of Molecular Pharmacology, National Cancer Institute, NIH, Bethesda, MD 20892.
W
W
Zhen
M K
MK
Evans
C M
CM
Haggerty
V A
VA
Bohr
D002460:Cell Line; D002874:Chromosome Mapping; D002945:Cisplatin / Q000633:toxicity*; D004247:DNA / Q000187:drug effects* / Q000235:genetics; D004249:DNA Damage; D004260:DNA Repair; D004275:DNA, Ribosomal / Q000187:drug effects / Q000235:genetics; D005199:Fanconi Anemia / Q000235:genetics*; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D005816:Genetic Complementation Test; D006801:Humans; D007700:Kinetics; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics; D013997:Time Factors; D014983:Xeroderma Pigmentosum / Q000235:genetics*
D004275:DNA, Ribosomal; D004247:DNA; D013762:Tetrahydrofolate Dehydrogenase; D002945:Cisplatin