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There are increasing reports showing the clinical significance of thep53polymorphism status in terms of the response to chemotherapy. We investigated whetherp53polymorphism and mutation were associated within vitrosensitivity to 5-fluorouracil (5-FU) in patients with colorectal cancer. Chemosensitivity to 5-FU was evaluated by the collagen gel droplet embedded culture drug sensitivity test. 5-FU sensitivity of tumor cells without inactivep53mutation in the arginine/arginine (Arg/Arg) variant was significantly higher than that of tumor cells with or without inactivep53mutation in other variants (p= 0.022), whereas the 5-FU sensitivity of tumor cells with inactivep53mutation in the Arg/Arg variant was significantly lower than that of tumor cells with or without inactivep53mutation in other variants (p= 0.002). In the Arg/Arg variant, apoptotic cells induced by 5-FU treatment in patients without inactivep53mutation were more markedly increased than those in patients with inactivep53mutation (p= 0.037). Bax and Bcl-2 protein expressions in tumor tissue treated with 5-FU were associated with both 5-FU sensitivity and the apoptotic cell count. Our data show that the Arg/Arg genotype without inactivep53mutation could be predictive of a more favorable response and the Arg/Arg genotype with inactivep53mutation a less favorable response to chemotherapy using 5-FU in CRC. The combination of thep53codon 72 polymorphism andp53mutation status is a potential predictive marker of sensitivity to 5-FU in CRC.