Summary
The in-vitro effects of hydroxyurea 5-FU and 5-FUdR have been extensively studied in experimental systems employing cell-line techniques. In this study we investigated the effects of these drugs on the levels of incorporation of labeled nucleosides into DNA in explants of intact rat colonic mucosa maintained in organ culture. The effects of the nucleoside transport inhibitors nitrobenzylthioinosine (NBMPR) and dipyridamole—which are modulators of antimetabolite cytotoxicity—on the incorporation of tritiated thymidine [(3H]TdR) into DNA were also studied. The incorporation of tritiated TdR into DNA was reduced by hydroxyurea but was not altered by either 5-FU or 5-FUdR. The levels of tritiated deoxyuridine were reduced by 5-FU and 5-FUdR in separate experiments; this is in keeping with thymidylate synthase inhibition. NBMPR and dipyridamole also reduced 3H-TdR incorporation into DNA. These results can be explained in terms of the known mechanisms of action of these drugs. This experimental model is therefore useful in assessing the effects of antimetabolites and nucleoside transport inhibitors in intact colonic mucosa.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Belt, J. A.; Noel, L. D. Nucleoside transport in Walker 256 rat carcinosarcoma and S49 mouse lymphoma cells. Biochem. J. 232:681–688; 1985.
Bowen, D.; Bailey, B. D.; Guernsey, L. A. Rate-limiting steps in the interactions of fluoropyrimidines and methotrexate. Eur. J. Cancer Clin. Oncol. 5:651–657; 1984.
Chabner, B. A. Pyrimidines antagonists. In: Chabner, B. A., ed. Pharmacologic principles of cancer treatment. Philadelphia: W. B. Saunders; 1982:183.
Eriksson, S.; Skog, S.; Tribukait, B., et al. Deoxyribonucleoside triphosphate metabolism and the mammalian cell cycle: effects of hydroxyurea on mutant and mild-type mouse S49 T-lymphoma cells. Exp. Cell. Res. 168:79–88; 1987.
Finney, K. J.; Ince, P., Appleton, D. R., et al. A comparison of crypt-cell proliferation in rat colonic mucosain vivo andin vitro. J. Anat. 149:177–188; 1984.
Gilbert, J. M. Adjuvant chemotherapy of the large bowel. Cancer Treat. Rev. 9:195–288; 1982.
Ince, P.; Appleton, D. R.; Finney, K. J., et al. Verapamil increases the sensitivity of primary human colorectal carcinoma tissue to vincristine. Br. J. Cancer 53:137–139; 1986.
Jackman, A. L.; Taylor, G. A.; Calvert, A. H., et al. Modulation of antimetabolite effects: effects of thymidine on the efficacy of quinazoline-based thymidylate synthetase inhibitor CB3717. Biochem. Pharmacol. 33:3269–3275; 1984.
Jackson, R. C.; Jackman, A. L.; Calvert, A. H. Biochemical effects of quinazoline inhibitor of thymidylate synthetase CB3717 on human lymphoblastoid cells. Biochem. Pharmacol. 32:3783–3790; 1983.
Kapuscinsky, J.; Skoczyslas, B. Simple and rapid fluorimetric method for DNA microassay. Anal. Biochem. 83:252–257; 1977.
Kufe, D. W.; Scott, P.; Fram, R., et al. Biologic effect of 5-fluoro-2′-deoxyuridine incorporation in L1210 deoxyribonucleic acid. Biochem. Pharmacol. 32:1337–1340; 1983.
Munro, H. N.; Fleck, A. The determination of nucleic acids. Methods Biochem. Anal. 14:113–176; 1966.
Paterson, A. R. P.; Lau, E. Y.; Dahlig, E., et al. A common basis for inhibition of nucleoside transport by dipyridamole and nitrobenzylthioinosine. Mol. Pharmacol. 18:40–44; 1980.
Paterson, A. R. P.; Kolassa, N.; Lynch, T. P., et al. In-vivo studies with an inhibitor of nucleoside transport nitrobenzylthioinosine-5-monophosphate. In: Tattersall, M. H. N.; Fox, R. M., eds. Nucleoside and cancer treatment: rational approach to antimetabolite selectivity and modulation. New York: Academic Press; 1981:84–95.
Patterson, M. K. Measurement of growth and viability of cells in culture. In: Jakoby, W. B., ed. Methods in enzymology, vol. LVIII. New York: Academic Press; 1979:141–152.
Peters, G. J.; Laurensse, E.; Leyva, A., et al. Sensitivity of human, murine and rat cells to 5-fluorouracil and 5-deoxy-5-fluorouridine in relation to drug metabolizing enzymes. Cancer Res. 46:20–28; 1986.
Plagemann, P. G. W.; Wohlhueter, R. M. Nucleoside transport in cultured mammalian cells. Multiple forms with different sensitivity to inhibition by nitrobenzylthioinosine or hypoxanthine. Biochim. Biophys. Acta 773:39–52; 1984.
Shi, M. M.; Young, J. D.3[H]Dipyridamole binding to nucleoside transporters from guinea-pig and rat lung. Biochem. J. 240:879–883; 1986.
Sinclair, W. K. Hydroxyurea revisited: a decade of clinical effects studies. Int. J. Radiat. Oncol. Biol. Phys. 7:631–637; 1981.
Snyder, R. D. Evaluation of putative inhibitors of DNA excision repair in cultured cells by the rapid nick translation assay. Mutat. Res. 173:279–286; 1986.
Sorger, T.; Germinario, R. J. A direct solubilisation method for the determination of DNA and protein in cultured fibroblast monolayers. Anal. Biochem. 131:254–256; 1983.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Moorghen, M., Ince, P., Finney, K.J. et al. Organ culture as a model for investigating the effects of antimetabolites and nucleoside transport inhibitors on rodent colonic mucosa. In Vitro Cell Dev Biol - Animal 27, 873–877 (1991). https://doi.org/10.1007/BF02630990
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02630990