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Sensitivity and Resistance to Therapy

Decitabine (5-Aza-2′-deoxycytidine) decreased DNA methylation and expression of MDR-1 gene in K562/ADM cells

Leukemia volume 14, pages 1915–1920 (2000)Cite this article

Abstract

Multidrug resistance (MDR) is a major problem in patients with hematological malignancies. Although drug-resistance is known to be induced by the expression of P-glycoprotein (P-gp) encoded by the MDR-1 gene, little is known about the mechanisms regulating this gene. Herein, we studied the DNA methylation patterns at the enhancer and repressor binding sites of the MDR-1 gene using the human erythroleukemia cell line K562 and its multidrug resistant derivative K562/ADM (adriamycin). Direct DNA sequence analysis demonstrated methylation to be present at the repressor site (minus 110 GC-box) of the MDR-1 gene in K562/ADM cells, but not in parental K562 cells. Methylation-specific PCR (MSP) analysis yielded similar results. Treatment of K562/ADM cells with 5-Aza-2′-deoxycytidine (decitabine; DAC), an inhibitor of DNA methyltransferase, caused demethylation of the repressor binding site of MDR-1 gene, as assessed by MSP, and also decreased P-gp expression, as assessed by flow cytometric and Northern blot analysis. Although it is generally accepted that DAC upregulates gene expression by demethylating the activator binding sites, our present results suggest that DAC induces down-regulation of P-gp expression as a result of demethylation at the repressor binding site in K562/ADM cells. In this regard, methylation-dependent regulation of the MDR-1 gene in K562/ADM cells is unique.

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References

  1. Riordan JR, Ling V . Genetic and biochemical characterization of multidrug resistance Pharmacol Ther 1985 28: 51–75

    Article  CAS  PubMed  Google Scholar 

  2. Ferro-Luzzi Ames G . The basis of multidrug resistance in mammalian cells: homology with bacterial transport Cell 1986 47: 323–324

    Article  CAS  PubMed  Google Scholar 

  3. Tsuruo T . Mechanisms of multidrug resistance and implications for therapy Jpn J Cancer Res 1988 79: 285–296

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Ueda K, Pastan I, Gottesman MM . Isolation and sequence of the promoter region of the human multidrug-resistance (P-glycoprotein) gene J Biol Chem 1987 262: 17432–17436

    CAS  PubMed  Google Scholar 

  5. Cornwell MM, Smith DE . SP1 activates the MDR1 promoter through one of two distinct G-rich regions that modulate promoter activity J Biol Chem 1993 268: 19505–19511

    CAS  PubMed  Google Scholar 

  6. McCoy C, Smith DE, Cornwell MM . 12-O-tetradecanoylphorbol-13-acetate activation of the MDR1 promoter is mediated by EGR1 Mol Cell Biol 1995 15: 6100–6108

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Holliday R, Grigg GW . DNA methylation and mutation Mutat Res 1993 285: 61–67

    Article  CAS  PubMed  Google Scholar 

  8. Bird A . The essentials of DNA methylation Cell 1992 70: 5–8

    Article  CAS  PubMed  Google Scholar 

  9. Adrian P . CpG-rich islands and the function of DNA methylation Nature 1986 321: 209–213

    Article  Google Scholar 

  10. Antequera F, Boyes J, Bird A . High levels of de novo methylation and altered chromatin structure at CpG islands in cell lines Cell 1990 62: 503–514

    Article  CAS  PubMed  Google Scholar 

  11. Herman JG, Latif F, Weng Y, Lerman MI, Zbar B, Liu S, Samid D, Duan DS, Gnarra JR, Linehan WM . Silencing of the VHL tumor-suppressor gene by DNA methylation in renal carcinoma Proc Natl Acad Sci USA 1994 91: 9700–9704

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Merlo A, Herman JG, Mao L, Lee DJ, Gabrielson E, Burger PC, Baylin SB, Sidransky D . 5′ CpG island methylation is associated with transcriptional silencing of the tumour suppressor p16/CDKN2/MTS1 in human cancers Nature Med 1995 1: 686–692

    Article  CAS  PubMed  Google Scholar 

  13. Herman JG, Jen J, Merlo A, Baylin SB . Hypermethylation associated inactivation indicates a tumor suppressor role for p15INK4B Cancer Res 1996 56: 722–727

    CAS  PubMed  Google Scholar 

  14. Graff JR, Herman JG, Lapidus RG, Chopra H, Xu R, Jarrard DF, Isaacs WB, Pitha PM, Davidson NE, Baylin SB . E-cadherin expression is silenced by DNA hypermethylation in human breast and prostate carcinomas Cancer Res 1995 55: 5195–5199

    CAS  PubMed  Google Scholar 

  15. Lubbert M, Mertelsmann R, Herrmann F . Cytosine methylation changes during normal hematopoiesis and in acute myeloid leukemia Leukemia 1997 11: (Suppl. 1) S12–S18

    Article  PubMed  Google Scholar 

  16. Batova A, Diccianni MB, Yu JC, Nobori T, Link MP, Pullen J, Yu AL . Frequent and selective methylation of p15 and deletion of both p15 and p16 in T-cell acute lymphoblastic leukemia Cancer Res 1997 57: 832–836

    CAS  PubMed  Google Scholar 

  17. Ottaviano YL, Issa JP, Parl FF, Smith HS, Baylin SB, Davidson NE . Methylation of the estrogen receptor gene CpG island marks loss of estrogen receptor expression in human breast cancer cells Cancer Res 1994 54: 2552–2555

    CAS  PubMed  Google Scholar 

  18. Haaf T . The effects of 5-azacytidine and 5-azadeoxycytidine on chromosome structure and function: implications for methylation-associated cellular processes Pharmacol Ther 1995 65: 19–46

    Article  CAS  PubMed  Google Scholar 

  19. Migeon BR . Insights into X chromosome inactivation from studies of species variation, DNA methylation and replication, and vice versa Genet Res 1990 56: 91–98

    Article  CAS  PubMed  Google Scholar 

  20. Li E, Beard C, Jaenisch R . Role for DNA methylation in genomic imprinting Nature 1993 366: 362–365

    CAS  PubMed  Google Scholar 

  21. Quesnel B, Guillerm G, Vereecque R, Wattel E, Preudhomme C, Bauters F, Vanrumbeke M, Fenaux P . Methylation of the p15(INK4b) gene in myelodysplastic syndromes is frequent and acquired during disease progression Blood 1998 91: 2985–2990

    CAS  PubMed  Google Scholar 

  22. Sakai T, Toguchida J, Ohtani N, Yandell DW, Rapaport JM, Dryja TP . Allele-specific hypermethylation of the retinoblastoma tumor-suppressor gene Am J Hum Genet 1991 48: 880–888

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Harrington MA, Jones PA, Imagawa M, Karin M . Cytosine methylation does not affect binding of transcription factor Sp1 Proc Natl Acad Sci USA 1998 85: 2066–2070

    Article  Google Scholar 

  24. Kantharidis P, El-Osta A, deSilva M, Wall DM, Hu XF, Slater A, Nadalin G, Parkin JD, Zalcberg JR . Altered methylation of the human MDR-1 promoter is associated with acquired multidrug resistance Clin Cancer Res 1997 3: 2025–2032

    CAS  PubMed  Google Scholar 

  25. Lau A, Gant TW, Cann AJ . The mechanism of trans-activation of the MDR1 gene by human T cell leukemia virus Biochem Biophys Res Commun 1998 249: 397–404

    Article  CAS  PubMed  Google Scholar 

  26. Fryxell KB, McGee SB, Simoneaux DK, Willman CL, Cornwell MM . Methylation analysis of the human multidrug resistance 1 gene in normal and leukemic hematopoietic cells Leukemia 1999 13: 910–917

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

The authors wish to thank Dr T Tsuruo (Institute of Molecular and Cellular Biosciences, The University of Tokyo) for his generous gift of K562/ADM cells and valuable technical advice.

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  1. The Third Department of Internal Medicine, Yamaguchi University School of Medicine, Minami-Kogushi 1-1-1, Yamaguchi, 755–8505, Japan

    T Ando, M Nishimura & Y Oka

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Ando, T., Nishimura, M. & Oka, Y. Decitabine (5-Aza-2′-deoxycytidine) decreased DNA methylation and expression of MDR-1 gene in K562/ADM cells. Leukemia 14, 1915–1920 (2000). https://doi.org/10.1038/sj.leu.2401914

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