Cell cycle regulation of membrane glucocorticoid receptor in CCRF-CEM human ALL cells: Correlation to apoptosis

Faustina N A Sackey, Cheryl S. Watson, Bahiru Gametchu

    Research output: Contribution to journalArticle

    3 Citations (Scopus)

    Abstract

    -The human leukemic cell line (CCRF-CEM) and a subline enriched for the plasma membrane-resident glucocorticoid receptor (mGR) were studied for the influence of the cell cycle on the expression and function of mGR. Three synchronization procedures (double thymidine, colcemid, and combined thymidine-colcemid blocks) were used. Fluorescent microscopy and flow cytometry simultaneously assessed antibody-tagged mGR and DNA. In addition, mGR was quantitated and characterized by immunoprecipitation and immunoblotting. Apoptosis was assayed by DNA fragmentation (TUNEL assay) and by cell survival (trypan blue exclusion). All synchronization procedures demonstrated that progression from DNA replication (S) to the second growth phase and mitosis (G2/M) leads to cells having the highest levels of mGR expression and being highly glucocorticoid sensitive in the apoptosis assays: 32 and 80% sensitivity of wild type and mGR-enriched cells, respectively, compared with 12 and 30% sensitivity in asynchronous cells. Therefore, mGR expression appears to be cell cycle regulated, with its highest expression at late S-G2/M, when the cells are most sensitive to the lymphocytolytic effects of glucocorticoids.

    Original languageEnglish (US)
    JournalAmerican Journal of Physiology
    Volume273
    Issue number3 PART 1
    StatePublished - 1997

    Fingerprint

    Glucocorticoid Receptors
    Cell Cycle
    Apoptosis
    Membranes
    Demecolcine
    Thymidine
    Glucocorticoids
    Trypan Blue
    G2 Phase
    In Situ Nick-End Labeling
    DNA Fragmentation
    DNA Replication
    Immunoprecipitation
    Immunoblotting
    Mitosis
    Microscopy
    Cell Survival
    Flow Cytometry
    Cell Membrane
    Cell Line

    Keywords

    • Acute lymphoblastic leukemic (all) cells
    • Lymphocytolysis
    • Steroids

    ASJC Scopus subject areas

    • Physiology (medical)

    Cite this

    Cell cycle regulation of membrane glucocorticoid receptor in CCRF-CEM human ALL cells : Correlation to apoptosis. / Sackey, Faustina N A; Watson, Cheryl S.; Gametchu, Bahiru.

    In: American Journal of Physiology, Vol. 273, No. 3 PART 1, 1997.

    Research output: Contribution to journalArticle

    Sackey, Faustina N A ; Watson, Cheryl S. ; Gametchu, Bahiru. / Cell cycle regulation of membrane glucocorticoid receptor in CCRF-CEM human ALL cells : Correlation to apoptosis. In: American Journal of Physiology. 1997 ; Vol. 273, No. 3 PART 1.
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