Mismatch repair proficiency and in vitro response to 5-fluorouracil.
OBJECTIVE
The DNA mismatch repair (MMR) system recognizes certain DNA adducts caused by alkylation damage in addition to its role in recognizing and directing repair of interstrand nucleotide mismatches and slippage mistakes at microsatellite sequences. Because defects in the MMR system can confer tolerance to acquired DNA damage and, by inference, the toxic effects of certain chemotherapeutic agents, we investigated the effect of 5-fluorouracil (5-FU) on colon cancer cell lines.


METHODS
We determined growth selection by cell enrichment assay and cloning efficiency after treatment with 5 micromol/L 5-FU, assayed nucleic 3H-5-FU incorporation, and analyzed the cell cycle by flow cytometry.


RESULTS
5-FU treatment provided a growth advantage for MMR-deficient cell lines, indicating a relative degree of tolerance to 5-FU by the MMR-deficient cell lines. Enhanced survival was statistically significant after 5 days of growth, and a 28-fold reduction in survival was noted in the MMR-proficient cells by clonagenic assays after 10 days of growth. Differences in nucleotide uptake of 5-FU did not account for the observed growth differences, and specific cell cycle checkpoint arrest was not detected.


CONCLUSIONS
Intact DNA MMR seems to recognize 5-FU incorporated into DNA but may do so in a different manner than other types of alkylation damage. Defective DNA MMR might be one mechanism for tumor resistance to 5-FU.
Carethers
J M
JM
Department of Medicine, University of California, USA. jcarethers@ucsd.edu
Chauhan
D P
DP
Fink
D
D
Nebel
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S
Bresalier
R S
RS
Howell
S B
SB
Boland
C R
CR
D000818:Animals; D000964:Antimetabolites, Antineoplastic / Q000378:metabolism / Q000494:pharmacology*; D020137:Base Pair Mismatch / Q000187:drug effects* / Q000502:physiology*; D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival / Q000502:physiology; D004260:DNA Repair / Q000187:drug effects* / Q000502:physiology*; D005472:Fluorouracil / Q000378:metabolism / Q000494:pharmacology*; D016195:G2 Phase / Q000502:physiology; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D008938:Mitosis / Q000502:physiology; D009696:Nucleic Acids / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000502:physiology
D000964:Antimetabolites, Antineoplastic; D009696:Nucleic Acids; D005472:Fluorouracil