Tumors with the microsatellite instability (MSI) phenotype appear to comprise a biologically and clinically distinct group of colorectal carcinomas (CRC). MSI+ has been associated with favorable prognosis; however, it is not clear whether this is because MSI+ tumors are inherently less aggressive or because they are more sensitive to chemotherapy. We investigated the prognostic and predictive significance of this molecular alteration along with its association with nodal burden in a large, population-based cohort of stage III CRC patients. Eight hundred seventy-six stage III CRC patients with long median follow-up (76 months) were included in the study. MSI status was determined by screening for deletions in the BAT-26 mononucleotide repeat. Systemic adjuvant fluoropyrimidine-based chemotherapy was delivered to 266 patients (30%). MSI+ was more common in tumors from female patients and tumors that originated in the proximal colon. It was predictive of excellent survival benefit from chemotherapy but was not associated with better prognosis for patients who did not receive treatment. Lower nodal burden was a prognostic factor for improved survival. MSI+ was associated with lower nodal burden in the overall group (P = 0.02, chi 2 test) but not for patients who received chemotherapy. In stage III CRC, MSI+ was not prognostic in nonadjuvant-treated patients, suggesting that the biological behavior of MSI+ tumors in the absence of chemotherapy is the same as MSI- tumors. Tumors with the MSI+ phenotype appear to be more sensitive to chemotherapy, as observed by improved survival for patients receiving this treatment. MSI along with other molecular markers could be used in the future for a more refined selection of CRC patients to receive fluoropyrimidine-based chemotherapy.