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PML-RARα and AML1–ETO translocations are rarely associated with methylation of the RARβ2 promoter

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Abstract

The acute promyelocytic leukemia-specific PML–RARα fusion protein is a dominant-negative transcriptional repressor of retinoic acid receptor (RAR) target genes, which recruits HDAC and corepressor proteins and inhibits coactivators. Another oncogenic transcription factor, AML1–ETO, was proposed to cause an HDAC-dependent repression of RAR target genes. The RAR target RARβ2 gene has been reported to be frequently silenced by hypermethylation in many types of cancer cells. We examined the methylation status of the RARβ2 and asked if demethylation could reverse ATRA resistance in ATRA-resistant PML–RARα and AML1–ETO-positive cells. PML–RARα positive NB4 and its ATRA-resistant subvariant MR2 and AML1–ETO expressing Kasumi-1 cells had heterozygous methylation of RARβ2. Although DNA methyltransferase inhibitor 5-aza-2′-deoxycytidine partially reversed RARβ2 CpG methylation in these cells, it did not significantly enhance ATRA-induced RARβ2 mRNA expression and induction of maturation. However, the histone acetylase inhibitor SAHA combined with ATRA significantly reactivated RARβ2 mRNA both in NB4 and MR2 cells with degradation of PML–RARα, which was associated with maturation. In contrast, SAHA did not affect AML1–ETO levels and failed to induce RARβ2 expression and maturation in Kasumi-1 cells. In primary AML samples, RARβ2 expression was uniformly low; however, no specific correlation was observed between the methylation of the RARβ2 gene and expression of the fusion proteins, PML–RARα, and AML1–ETO. These results demonstrate that oncogenic PML–RARα and AML1–ETO translocations are rarely associated with RARβ2 promoter methylation in primary AML samples.

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Acknowledgements

We thank Betty Notzon for manuscript review, Nobuko Tanaka, Hiroaki Miyajima, and Akemi Koyanagi for technical assistance, and Rosemarie Lauzon for help in the preparation of the manuscript. This study is supported in part by grants from the National Cancer Institute (PO1 CA55164, PO1 CA49639, RO1 CA89346, and CA16672) to M.A. and the Leukemia and Lymphoma Society (6097-02) to M.K.

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Authors and Affiliations

  1. Section of Molecular Hematology and Therapy, Department of Blood and Marrow Transplantation, The University of Texas M. D. Anderson Cancer Center, 1400 Holcombe Boulevard, Unit 448, Houston, TX, 77030, USA

    Yoko Tabe, Marina Konopleva, Rooha Contractor, Vivian Ruvolo & Michael Andreeff

  2. Department of Leukemia, The University of Texas M.D. Anderson Cancer Center, Houston, TX, 77030, USA

    Yutaka Kondo, Jean-Pierre J. Issa & Michael Andreeff

  3. Department of Clinical Pathology, Juntendo University School of Medicine, Tokyo, Japan

    Yoko Tabe, Linhua Jin, Kazunori Miyake & Noriko Miyake

  4. Department of Biochemistry, Juntendo University School of Medicine, Tokyo, Japan

    Yuko Tsutsumi-Ishii & Isao Nagaoka

  5. Department of Transfusion Medicine and Stem Cell Regulation, Juntendo University School of Medicine, Tokyo, Japan

    Akimichi Ohsaka

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  1. Yoko Tabe
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Correspondence to Michael Andreeff.

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Tabe, Y., Konopleva, M., Kondo, Y. et al. PML-RARα and AML1–ETO translocations are rarely associated with methylation of the RARβ2 promoter. Ann Hematol 85, 689–704 (2006). https://doi.org/10.1007/s00277-006-0148-7

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