Summary
Differentiation therapy provides an alternative treatment of cancer that overcomes the undesirable effects of classical chemotherapy, i.e. cytotoxicity and resistance to drugs. This new approach to cancer therapy focuses on the development of specific agents designed to selectively engage the process of terminal differentiation, leading to the elimination of tumorigenic cells and recovery of normal cell homeostasis. A series of new anti-cancer pyrimidine acyclonucleoside-like compounds were designed and synthesized by structural modifications of 5-fluorouracil, a drug which causes considerable cell toxicity and morbidity, and we evaluated their applicability for differentiation therapy in human rhabdomyosarcoma cells. We tested the pyrimidine derivative GR-891, (RS)-1-{[3-(2-hydroxyethoxy)-1-isopropoxy]propyl}-5-fluorouracil, an active drug which shows low toxicity in vivo and releases acrolein which is an aldehyde with anti-tumour activity. Both GR-891 and 5-fluorouracil caused time- and dose-dependent growth inhibition in vitro; however, GR-891 showed no cytotoxicity at low doses (22.5 μmol l–1 and 45 μmol l–1) and induced terminal myogenic differentiation in RD cells (a rhabdomyosarcoma cell line) treated for 6 days. Changes in morphological features and in protein organization indicated re-entry in the pathway of muscular maturation. Moreover, GR-891 increased adhesion capability mediated by the expression of fibronectin, and did not induce overexpression of P-glycoprotein, the mdr1 gene product, implicated in multidrug resistance. New acyclonucleoside-like compounds such as GR-891 have important potential advantages over 5-fluorouracil because of their lower toxicity and their ability to induce myogenic differentiation in rhabdomyosarcoma cells. Our results suggest that this drug may be useful for differentiation therapy in this type of tumour.
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References
Aguanno, S., Bouche, M., Adamo, S. & Molinaro, M. (1990). 12-O-Tetradecanoylphorbol-13-acetate-induced differentiation of a human rhabdomyosarcoma cell line. Cancer Res 50: 3377–3382.
Beere, H. M. & Hickman, J. A. (1993). Differentiation: a suitable strategy for cancer chemotherapy?. Anticancer Drug Dev 8: 299–322.
Borden, E. C., Lotan, R., Levens, D., Young, C. W. & Waxman, S. (1993). Differentiation therapy of cancer: laboratory and clinical investigations. Cancer Res 53: 4109–4115.
Burridge, K. & Feramisco, J. R. (1981). Non-muscle α-actinins are calcium-sensitive actin binding proteins. Nature 294: 565–567.
Campos, J., Pineda, M. J., Gómez, J. A., Entrena, A., Trujillo, M. A., Gallo, M. A. & Espinosa, A. (1996). 5-fluorouracil derivatives. 1. Acyclonucleosides through a tin (IV) chloride-mediated regiospecific ring opening of alkoxy-1,4-diheteroepanes. Tetrahedron 52: 8907–8924.
Capetanaki, Y. G., Ngai, J. & Lazarides, E. (1984). Characterization and regulation in the expression of a gene coding for the intermediate filament desmin. Proc Natl Acad Sci USA 81: 6909–6913.
Carter, R. L., McCarthy, K. P., Machin, L. G., Jameson, C. F., Philip, E. R. & Pinkerton, C. R. (1989). Expression of desmin and myoglobin in rhabdomyosarcomas and developing skeletal muscle. Histopathology 1: 585–595.
Chen, Z. X. & Breitman, T. R. (1994). Tributyrin: a prodrug of butyric acid for potential clinical application in differentiation therapy. Cancer Res 54: 3494–3499.
Crouch, G. D. & Helman, L. J. (1991). All-trans-retinoic acid inhibits the growth of human rhabdomyosarcoma cell lines. Cancer Res 51: 4882–4887.
Crouch, G. D., Kalebic, T., Tsokos, M. & Helman, L. J. (1993). Ara-C treatment leads to differentiation and reverses the transformed phenotype in a human rhabdomyosarcoma cell line. Exp Cell Res 204: 210–216.
D’Amore, E. S. G., Tollot, M., Stracca-Pansa, V., Menegon, A., Mell, S. & Ninfo, V. (1994). Therapy associated differentiation in rhabdomyosarcomas. Modern Pathol 7: 69–75.
De Giovanni, C., Lollini, P. L., Dolcetti, R., Landuzzi, L., Nicpletti, G., D’Andrea, E., Scotland, K. & Nanni, P. (1993). Uncoupling of growth inhibition and differentiation in dexamethasone-treated human rhabdomyosarcoma cells. Br J Cancer 67: 674–679.
Degos, L. (1995). A new concept: treatment using differentiation of the malignant cell in man. Bull Acad Natl Med 179: 1689–1700.
Enomoto, M. I., Boettiger, D. & Menko, A. S. (1993). Alpha 5 integrin is a critical component of adhesion plaques in myogenesis. Dev Biol 155: 180–197.
Eyden, B. P. (1993). Brief review of the fibronexus and its significance for myofibroblastic differentiation and tumor diagnosis. Ultrastruct Pathol 17: 611–622.
Farquhar, D., Chen, R. & Khan, S. (1995). 5′-[4-(Pivaloyloxy)-1,3,2-dioxaphosphorinan-2-yl]-2′-deoxy-5-fluorouridine: a membrane-permeating prodrug of 5-fluoro-2′-deoxyuridylic acid (FdUMP). J Med Chem 38: 488–495.
Fuqua, S. A., Fitzgerald, S. D. & McGuire, W. L. (1990). A simple polymerase chain reaction method for detection and cloning of low-abundance transcripts. Biotechniques 9: 206–211.
Gabbert, H. E., Gerharz, C. D., Biesalsk, H. K., Engers, R. & Luley, C. (1988). Terminal differentiation and growth inhibition of a rat rhabdomyosarcoma cell line (BA-HAN-1C) in vitro after exposure to retinoic acid. Cancer Res 48: 5264–5269.
Germann, U. A. (1996). P-glycoprotein: a mediator of multidrug resistance in tumour cells. Eur J Cancer 32: 927–944.
Gómez, J. A., Campos, J., Marchal, J. A., Trujillo, M. A., Melguizo, C., Prados, J., Gallo, M. A., Aránega, A. & Espinosa, A. (1997). Chemical modifications on the acyclic moiety of 3-(2-hydroxyethoxy)-1-alkoxypropyl nucleobases. 2. Differentiation and growth inhibition in rhabdomyosarcoma cells after exposure to a novel 5-fluorouracil acyclonucleoside. Tetrahedron 53: 7319–7334.
Gundersen, G. G., Khawaja, S. & Bulinski, J. C. (1989). Generation of a stable, post-translationally modified microtubule array is an early event in myogenic differentiation. J Cell Biol 109: 2275–2288.
Hazelton, B. J., Houghton, J. A., Parham, D. M., Douglass, E. L., Torrance, P. M., Holt, H. & Houghton, P. J. (1987). Characterization of cell lines derived from xenographs of childhood rhabdomyosarcoma. Cancer Res 47: 4501–4507.
Herrera, A., Royal, A. & Babai, F. (1995). Correlation between cell differentiation, types of invasion, and hematogenous metastasis in experimental rhabdomyosarcomas. Exp Mol Pathol 63: 1–15.
Langbein, L., Kosmehl, H., Katenkamp, D., Neupert, G. & Stiller, K. J. (1990). Experimentally induced murine rhabdomyosarcomas – correlation between cellular contacts, matrix formation and cellular differentiation. Differentiation 44: 185–196.
Le Moigne, A., Martelly, I., Barlovatz, G., Franquinet, R., Aamiri, A., Frisdal, E., Bassaglia, Y., Moraczewski, G. & Gautron, J. (1990). Characterization of myogenesis from adult satellite cells cultured in vitro. Int J Dev Biol 34: 171–180.
Lesuffleur, T., Kornowski, A., Augeron, C., Dussaulx, E., Barbat, A., Laboisse, C. & Zweibaum, A. (1991a). Increased growth adaptability to 5-fluorouracil and methotrexate of HT-29 subpopulations selected for their commitment to differentiation. Int J Cancer 49: 731–737.
Lesuffleur, T., Kornowski, A., Luccioni, C., Muleris, M., Barbat, A., Beaumatin, J., Dussaulx, E., Dutrillaux, B. & Zweibaum, A. (1991b). Adaptation to 5-fluorouracil of the heterogeneous human colon tumor cell line HT-29 results in the selection of cells committed to differentiation. Int J Cancer 49: 721–730.
Lollini, P. L., De Giovanni, C., Del Re, B., Landuzzi, L., Nicoletti, G. & Prodi, G. (1989). Myogenic differentiation of human rhabdomyosarcoma cells induced in vitro by antineoplastic drugs. Cancer Res 49: 3631–3636.
MacAllister, R. M., Melnyk, J., Finklestein, J. Z., Adams, E. C. & Gardner, M. B. (1969). Cultivation in vitro of cells derived from a human rhabdomyosarcoma. Cancer 24: 520–526.
Maniatis, T., Fristch, E. & Sambrook, J. (1982). Molecular cloning. In Cold Spring Harbor, A Laboratory Manual, Cold Spring Harbor Laboratory New York
Marchal, J. A., Prados, J., Melguizo, C., Fernández, J. E., Vélez, C., Alvarez, L. & Aránega, A. (1997). Actinomycin D treatment leads to differentiation and inhibits proliferation in rhabdomyosarcoma cells. J Lab Clin Med 130: 42–50.
Melguizo, C., Prados, J., Aneiros, J., Fernández, J. E., Vélez, C. & Aránega, A. (1995). Differentiation of a human rhabdomyosarcoma cell line after antineoplastic drug treatment. J Pathol 175: 23–29.
Melguizo, C., Prados, J., Marchal, J. A., Aránega, A. E., Alvarez, L. & Aránega, A. (1996). Low concentrations of actinomycin D potentially cause therapeutic differentiation in human rhabdomyosarcoma cell line RD. Path. Res Pract 192: 188–194.
Melo, F., Carey, D. J. & Brandan, E. (1996). Extracellular matrix is required for skeletal muscle differentiation but not myogenin expression. J Cell Biochem 62: 227–239.
Nootan, K. E., Beck, C., Holzmayer, T. A., Chin, J. E., Wunder, J. S., Andrulis, I. L., Gazdar, A. F., Willman, C. L., Griffith, B., Von Hoff, D. D. & Roninson, I. B. (1990). Quantitative analysis of MDR1 (multidrug resistance) gene expression in human tumors by polymerase chain reaction. Proc Natl Acad Sci USA 87: 7160–7164.
Prados, J., Melguizo, C., Fernández, J. E., Aránega, A. E., Alvarez, L. & Aránega, A. (1993). Actin, tropomyosin and α-actinin as markers of differentiation in human rhabdomyosarcoma cell lines induced with dimethyl sulfoxide. Cell Mol Biol 39: 525–536.
Prados, J., Melguizo, C., Fernández, J. E., Aránega, A. E., Alvarez, L. & Aránega, A. (1996). Inverse expression of mdr1 and c-myc genes in a rhabdomyosarcoma cell line resistant to actinomycin D. J Pathol 180: 85–89.
Prados, J., Melguizo, C., Marchal, J. A., Vélez, C., Alvarez, L. & Aránega, A. (1998). Therapeutic differentiation in a human rhabdomyosarcoma cell line selected for resistance to actinomycin D. Int J Cancer 75: 379–383.
Sartorelli, A. (1985). Malignant cell differentiation as a potential therapeutic approach. Br J Cancer 52: 293–302.
Skach, W. R. (1996). Transmembrane orientation and topogenesis of the third and fourth membrane-spanning regions of human P-glycoprotein (MDR1). Cancer Res 54: 3202–3209.
Vogel, W., Kosmehl, H., Katenkamp, D. & Langbein, L. (1991). Differentiation dependent matrix formation (fibronectin and laminin) in an experimental murine rhabdomyosarcoma model. Acta Histochem 90: 181–188.
Volk, T., Fessler, L. I. & Fessler, J. H. (1990). A role for integrin in the formation of sarcomeric cytoarchitecture. Cell 63: 525–536.
Waxman, S., Scher, B. M., Hellinger, N. & Scher, W. (1990). Combination cytotoxic-differentiation therapy of mouse erythroleukemia cells with 5-fluorouracil and hexamethylene bisacetamide. Cancer Res 50: 3878–3887.
Waxman, S., Huang, Y., Scher, B. M. & Scher, M. (1992). Enhancement of differentiation and cytotoxicity of leukemia cells by combinations of fluorinated pyrimidines and differentiation inducers: development of DNA double-strand breaks. Biomed Pharmacother 46: 183–192.
Wijnaendts, L. C. D., Van Der Linden, J. C., Van Unnik, A. J. M., Delemarre, J. F. M., Voute, P. A. & Meijer, C. J. L. (1994). The expression pattern of contractile and intermediate filament proteins in developing skeletal muscle and rhabdomyosarcoma of childhood: diagnostic and prognostic utility. J Pathol 174: 283–292.
Zupi, G., Marangolo, M., Arancia, G., Greco, C., Laudonio, N., Iosi, F., Formisano, G. & Malorni, W. (1988). Modulation of the cytotoxic effect of 5-fluorouracil by N-methylformamide on a human colon carcinoma cell line. Cancer Res 48: 6193–6200.
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Marchal, J., Prados, J., Melguizo, C. et al. GR-891: a novel 5-fluorouracil acyclonucleoside prodrug for differentiation therapy in rhabdomyosarcoma cells. Br J Cancer 79, 807–813 (1999). https://doi.org/10.1038/sj.bjc.6690129
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DOI: https://doi.org/10.1038/sj.bjc.6690129
