The effect of activation of wild-type p53 function on fluoropyrimidine-mediated radiosensitization.
OBJECTIVE
We have hypothesized that fluoropyrimidine-mediated (FdUrd) radiosensitization occurs in cells that inappropriately enter S phase in the presence of drug, resulting in defective repair of radiation-induced DNA damage (14). This model would predict that prevention of entry into S phase would abrogate sensitization produced by FdUrd. We wished to test this prediction by blocking S phase entry of HT29 human colon cancer cells.


METHODS
We used HT29 cells that had been transduced with a murine temperature-sensitive p53 (ts29G) and, as a control, HT29 cells transduced with a neomycin plasmid (HT29neo). The murine temperature-sensitive p53 demonstrates wild-type p53 function when cells are incubated at the permissive temperature (32 degrees C) and mutant p53 function at the nonpermissive temperature (38 degrees C). We determined the effect of wt p53 function on FdUrd-mediated radiosensitization and cell cycle progression.


RESULTS
Incubation of ts29G cells at the permissive temperature (32 degrees C) activated p21 and blocked entry of cells into S phase. Whereas FdUrd greatly increased the radiosensitivity of HT29neo cells and ts29G cells incubated at 38 degrees C, FdUrd had no effect on the radiation sensitivity of ts29G cells incubated at the permissive temperature (32 degrees C).


CONCLUSIONS
These findings are consistent with our hypothesis that FdUrd-mediated radiosensitization requires progression into S phase. It is possible that the heterogeneity of clinical responses seen after combined therapy with fluoropyrimidines and radiation is explained, in part, by differences among tumor cells in the control of S phase progression.
Naida
J D
JD
Department of Radiation Oncology, The University of Michigan Medical Center, Ann Arbor 48109-0010, USA.
Davis
M A
MA
Lawrence
T S
TS
D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology*; D005467:Floxuridine / Q000494:pharmacology*; D019073:HT29 Cells / Q000187:drug effects / Q000528:radiation effects; D006801:Humans; D009363:Neoplasm Proteins / Q000378:metabolism*; D011838:Radiation-Sensitizing Agents / Q000494:pharmacology*; D016196:S Phase / Q000187:drug effects* / Q000528:radiation effects; D013696:Temperature; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*
D000964:Antimetabolites, Antineoplastic; D009363:Neoplasm Proteins; D011838:Radiation-Sensitizing Agents; D016159:Tumor Suppressor Protein p53; D005467:Floxuridine