Abstract
Chemotherapy is frequently limited by the development of multidrug resistance, a major cause of which is activation of the P-glycoprotein-encodingMDR1 gene. We have previously developed a P-glycoprotein-expressing multidrug resistant subline (HL60/E8) from the non-P-glycoprotein-expressing human HL60 promyelocytic leukemia cell line. A possible cause ofMDR1 silencing in HL60 cells is methylation of the promoter proximal region, thus demethylation occurring as a result of drug treatment may be responsible forMDR1 activation in the multidrug resistant subline. Using the bisulphite genomic sequencing technique we demonstrated that HL60 DNA is methylated at multiple sites within two distinct areas, one upstream and one downstream of the transcription start point. Only a single site in each area was methylated in all strands examined, with the remaining adjacent sites showing partial methylation. In contrast, DNA from the multidrug resistant HL60/E8 subline was unmethylated at essentially all sites in both areas. Thus the development of the P-glycoprotein-expressing multidrug resistant subline was associated with demethylation of theMDR1 5′ region.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Literature Cited
Lehnert, M. (1996).Eur. J. Cancer 32A:912–920.
Gottesman, M.M., and Pastan, I. (1993).Ann. Rev. Biochem. 62:385–427.
Kane, S.E. (1996).Adv. Drug Res. 28:181–252.
Su, G.M.I., Davey, M.W., Davey, R.A., and Kidman, A.D. (1994).Brit. J. Haemat. 88:566–574.
Noonan, 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., and Roninson, I.B. (1990).Proc. Natl. Acad. Sci. U.S.A.,87:7160–7164.
Rischin, D., and Ling, V. (1993) In: Leukemia:Advances in Research and Treatment. (Freireich, E.J., and Kantarjian, H., Editors) pp. 269–294 Kluwer Academic Publishers, London.
Bradley, G., Naik, M., and Ling, V. (1989).Cancer Res. 49:2790–2796.
Shemer, R., and Razin, A. (1996) In:Epigenetic Mechanisms of Gene Regulation (Russo, V.E.A., Martienssen, R.A. and Riggs, A.D., Editors), pp 215–229 Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
Riggs, A.D., and Porter, T.N. (1996). In:Epigenetic Mechanisms of Gene Regulation (Russo, V.E.A., Martiensen, R.A., and Riggs, A.D., Editors), pp. 231–248, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
Antequera, F., Boyes, J., and Bird, A. (1990).Cell,62:503–514.
Madden, M.J., Morrow, C.S., Nakagawa, M., Goldsmith, M.E., Fairchild, C.R., and Cowan, K.H. (1993).J. Biol. Chem. 268:8290–8297.
Clark, S.J., Harrison, J., Paul, C.L., and Frommer, M. (1994).Nucleic Acids Res. 22:2990–2997.
Sambrook, J., Fritsch, E.F., and Maniatis, T. (1989). Molecular Cloning. A Laboratory Manual. 2nd Ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
Pastan, I., Gottesman, M.M., Ueda, K., Lovelace, E., Rutherford, A.V., and Willingham, M.C. (1988).Proc. Natl. Acad. Sci. U.S.A. 85:4486–4490.
Shen, D.W., Fojo, A., Chin, J.E., Roninson, I.B., Richert, N., Pastan, I., and Gottesman, M.M. (1986).Science 232:643–645.
Fuqua, S.A.W., Moretti-Rojas, I.M., Schneider, S.L., and McGuire, W.L. (1987).Cancer Res. 47:2103–2106.
Nelson, M., Raschke, E., and McClelland, M. (1993).Nucl. Acids Res. 21:3139–3154.
Gardiner-Garden, M., and Frommer, M. (1987).J. Mol. Biol. 196:261–282.
Lincke, C.R., Smitt, J.J.M., van der Velde-Koerts, T., and Borst P. (1991).J. Biol. Chem. 266:5303–5310.
Cornwell, M.M., and Smith, D.E. (1993).J. Biol. Chem. 268:19505–19511.
Hoeller, M., Westin, G., Jiricny, J. and Schaffner, W. (1988).Genes Dev. 2:1127–1135.
Combates, N.J., Rzepka, R.W., Chen, Y.-N.P., and Cohen, D. (1994).J. Biol. Chem. 269:29715–29719.
Ogura, M., Takatori, T., Tsuruo, T. (1992).Nucleic Acids Res. 20: 5811–5817.
Goldsmith, M.E., Madden, M.J., Morrow, C.S., and Cowan, K.H. (1993).J. Biol. Chem. 268:5856–5860.
Asakuno, K., Kohno, K., Uchiumi, T., Kubo, T., Sato, S., Isono, M., and Kuwano, M. (1994).Biochem. Biophys. Res. Commun. 199:1428–1435.
Boyes, J., and Bird, A. (1991).Cell 64:1123–1134.
Nan, X., Campoy, F.J., and Bird, A. (1997).Cell 88:471–481
Issa, J.P.J., Zehnbauer, B.A., Kaufman, S.H., Biel, M.A., and Baylin, S.B. (1997).Cancer Res. 57:1768–1681.
Stirzaker, C., Millar, D., Paul, C., Warnecke, P., Harrison, J. Vincent, P., Frommer, M. and Clark, S. (1997),Cancer Res. (in press).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Desiderato, L., Davey, M.W. & Piper, A.A. Demethylation of the humanMDR1 5′ region accompanies activation of P-glycoprotein expression in a HL60 multidrug resistant subline. Somat Cell Mol Genet 23, 391–400 (1997). https://doi.org/10.1007/BF02673749
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02673749