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MDS And Clonal Leukemic Differentiation

Methylation status of the p15INK4B gene in hematopoietic progenitors and peripheral blood cells in myelodysplastic syndromes

Leukemia volume 14, pages 586–593 (2000)Cite this article

Abstract

We previously reported that the hypermethylation of the p15INK4B gene promoter was frequently observed in myelodysplastic syndromes (MDS), and that it may be associated with disease progression. An unanswered question is whether p15INK4B gene methylation is restricted to undifferentiated blastic cells, or whether differentiated cells such as granulocytes or erythrocytes of MDS origin also harbor this epigenetic alteration. In this study, we analyzed the methylation status of the p15INK4B gene in MDS by the methylation-specific PCR (MSP) method, which is more sensitive than Southern blotting. The bone marrow mononuclear cells (BM-MNCs) of 23 MDS patients were analyzed, and six of them showed p15INK4B methylation. Progenitor assay with methylcellulose medium was also performed in all patients. In two of the six patients with p15INK4B-methylated BM-MNCs, erythroid and/or non-erythroid colonies formed were subjected to molecular analysis. Colonies with and without p15INK4B methylation were detected in both patients. Furthermore, X-chromosome inactivation (XCI) pattern of each colony was simultaneously determined by MSP-based human androgen receptor gene analysis (HUMARA-MSP), and all p15INK4B-methylated colonies showed the same XCI pattern, which was dominant among the colonies, while p15INK4B-unmethylated colonies showed both patterns of XCI, in each of the two patients. We then examined the methylation status of the p15INK4B gene of granulocyte (PB-PMN) fractions from 10 patients with available peripheral blood cells. In all four patients with p15INK4B-methylated BM-MNCs, their PB-PMNs showed p15INK4B methylation. These results suggest that p15INK4Bmethylation in hematopoietic cells in MDS patients is restricted to the MDS clone but not necessarily to blast cells.

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Acknowledgements

We thank Dr Suzuki and Dr Nakahara for suggestions on the experimental procedures, and Ms Hatano and Ms Kondo for technical assistance. This study was supported in part by the Ministry of Health and Welfare's Research Grant for Specific Disease, Japan.

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Authors and Affiliations

  1. The First Department of Internal Medicine, Nagoya University School of Medicine, Nagoya, Japan

    E Aoki, T Uchida, H Ohashi, H Nagai, T Kinoshita, H Saito & T Murate

  2. Toyota Memorial Hospital, Toyota, Japan

    T Murase

  3. Gifu Prefectural Tajimi Hospital, Tajimi, Japan

    A Ichikawa

  4. Chukyo Hospital, Nagoya, Japan

    K Yamao

  5. The Department of Internal Medicine, Tokai University School of Medicine, Isehara, Japan

    T Hotta

  6. Nagoya University School of Health Science, Nagoya, Japan

    T Murate

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Aoki, E., Uchida, T., Ohashi, H. et al. Methylation status of the p15INK4B gene in hematopoietic progenitors and peripheral blood cells in myelodysplastic syndromes. Leukemia 14, 586–593 (2000). https://doi.org/10.1038/sj.leu.2401719

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