Epigenetic alteration by DNA-demethylating treatment restores apoptotic response to glucocorticoids in dexamethasone-resistant human malignant lymphoid cells

Aaron L. Miller, Chuandong Geng, Georgiy Golovko, Meenakshi Sharma, Jason R. Schwartz, Jiabin Yan, Lawrence Sowers, William R. Widger, Yuriy Fofanov, Wayne V. Vedeckis, E. B. Thompson

    Research output: Contribution to journalArticlepeer-review

    9 Scopus citations

    Abstract

    Background: Glucocorticoids (GCs) are often included in the therapy of lymphoid malignancies because they kill several types of malignant lymphoid cells. GCs activate the glucocorticoid receptor (GR), to regulate a complex genetic network, culminating in apoptosis. Normal lymphoblasts and many lymphoid malignancies are sensitive to GC-driven apoptosis. Resistance to GCs can be a significant clinical problem, however, and correlates with resistance to several other major chemotherapeutic agents.Methods: We analyzed the effect of treatment with the cytosine analogue 5 aza-2' deoxycytidine (AZA) on GC resistance in two acute lymphoblastic leukemia (T or pre-T ALL) cell lines- CEM and Molt-4- and a (B-cell) myeloma cell line, RPMI 8226. Methods employed included tissue culture, flow cytometry, and assays for clonogenicity, cytosine extension, immunochemical identification of proteins, and gene transactivation. High throughput DNA sequencing was used to confirm DNA methylation status.Conclusions: Treatment of these cells with AZA resulted in altered DNA methylation and restored GC-evoked apoptosis in all 3 cell lines. In CEM cells the altered epigenetic state resulted in site-specific phosphorylation of the GR, increased GR potency, and GC-driven induction of the GR from promoters that lie in CpG islands. In RPMI 8226 cells, expression of relevant coregulators of GR function was altered. Activation of p38 mitogen-activated protein kinase (MAPK), which is central to a feed-forward mechanism of site-specific GR phosphorylation and ultimately, apoptosis, occurred in all 3 cell lines. These data show that in certain malignant hematologic B- and T-cell types, epigenetically controlled GC resistance can be reversed by cell exposure to a compound that causes DNA demethylation. The results encourage studies of application to in vivo systems, looking towards eventual clinical applications.

    Original languageEnglish (US)
    Article number35
    JournalCancer Cell International
    Volume14
    Issue number1
    DOIs
    StatePublished - Apr 23 2014

    Keywords

    • 5 Aza-2' deoxycytidine
    • Apoptosis
    • CEM
    • Dexamethasone
    • Epigenetic
    • Glucocorticoid
    • Glucocorticoid receptor
    • High throughput sequencing
    • Leukemia
    • Lymphoid
    • Methylation
    • Molt-4
    • Myeloma
    • p38
    • Phosphorylation
    • RPMI 8226

    ASJC Scopus subject areas

    • Cancer Research
    • Oncology
    • Genetics

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