Mesalazine improves replication fidelity in cultured colorectal cells.
Epidemiologic studies indicate that mesalazine has chemopreventive effects in inflammatory bowel disease-associated colorectal cancer. Most of our general understanding of chemoprevention in colorectal cancer is, however, derived from aspirin, which is structurally similar to mesalazine. Herein we determined the influence of aspirin and mesalazine on replication fidelity in cultured colorectal cells. Flow cytometry was used for quantitation of mutation rates at a (CA)13 microsatellite in HCT116 cells (mismatch repair deficient) and HCT116+chr3 cells (mismatch repair proficient) that had been stably transfected with pIREShyg2-EGFP/CA13, an enhanced green fluorescence protein-based plasmid, and cultured in the absence or presence of various concentrations of aspirin or mesalazine. Aspirin at doses above 1.25 mmol/L markedly reduced cell growth. Mesalazine doses up to 5.0 mmol/L had no such effect. The mutation rate in mismatch repair-deficient HCT116 cells was 6.8 x 10(-4) +/- 9.0 x 10(-5). In aspirin-treated cultures the mutation rate was 8.2 x 10(-4) +/- 1.3 x 10(-4) (121% of control). Instead, mesalazine lowered the mutation rate in a dose-dependent fashion (5.5 x 10(-4) +/- 1.1 x 10(-4); 81% of control). The effects of mesalazine were most significant in the M1 fraction (P < 0.0001), which represents a mutant population immediate after the polymerase error and were confirmed in mismatch repair-proficient HCT116+chr3 cells. Our data indicate that mesalazine reduces frameshift mutations at a (CA)13 microsatellite in cultured colorectal cells independent of mismatch repair proficiency. This finding suggests that mesalazine improves replication fidelity, an effect that may be active in reducing mutations independent of its anti-inflammatory properties.
Gasche
Christoph
C
Department of Medicine 4, Division of Gastroenterology and Hepatology, Medical University Vienna, Vienna, Austria.
Goel
Ajay
A
Natarajan
Loki
L
Boland
C Richard
CR
D001241:Aspirin / Q000494:pharmacology; D020137:Base Pair Mismatch; D002470:Cell Survival / Q000187:drug effects; D015179:Colorectal Neoplasms / Q000235:genetics* / Q000473:pathology* / Q000517:prevention & control; D004260:DNA Repair; D004261:DNA Replication / Q000187:drug effects*; D005434:Flow Cytometry; D016368:Frameshift Mutation / Q000187:drug effects; D045325:HCT116 Cells; D006801:Humans; D019804:Mesalamine / Q000494:pharmacology*
D019804:Mesalamine; D001241:Aspirin