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Isolation of mutant adrenocortical tumor cells resistant to cyclic nucleotides

B P Schimmer, J Tsao, M Knapp

    Molecular and Cellular Endocrinology
    |August 1, 1977
    PubMed

    Insights

    This study reveals a defect in cyclic AMP signaling in a mutant cell line, impacting adrenal steroidogenesis, cell growth, and morphology. These findings suggest cyclic AMP mediates ACTH

    Area of Science:

    • Endocrinology
    • Molecular Biology
    • Cell Biology

    Background:

    • Cyclic nucleotides play a role in regulating adrenal steroidogenesis.
    • Understanding the precise mechanisms of cyclic nucleotide action is crucial for adrenal function.

    Purpose of the Study:

    • To investigate the role of cyclic nucleotides in adrenal steroidogenesis using a somatic cell genetic approach.
    • To identify the specific molecular defects responsible for altered steroidogenesis in response to cyclic AMP.

    Main Methods:

    • Utilized cultured mouse adrenocortical tumor cells (Clone Y1).
    • Administered 8-Bromoadenosine 3',5'-monophosphate (8BrcAMP) to stimulate steroidogenesis and observe effects on cell growth and morphology.
    • Mutagenized Y1 cells and selected for resistance to 8BrcAMP-induced growth inhibition (8BrcAMPr-1 cells).
    • Assessed steroidogenic responses, adenylate cyclase, and phosphodiesterase activities in wild-type and mutant cells.

    Main Results:

    • 8BrcAMP stimulated steroidogenesis but inhibited Y1 cell growth and altered morphology.
    • A mutant cell line (8BrcAMPr-1) exhibited resistance to 8BrcAMP's effects on growth and morphology.
    • 8BrcAMPr-1 cells showed diminished steroidogenic responses to cyclic nucleotides and ACTH.
    • Adenylate cyclase remained responsive to ACTH in mutant cells, while phosphodiesterase activity was not elevated.

    Conclusions:

    • The 8BrcAMPr-1 cell line harbors a defect common to cyclic AMP action on growth, morphology, and steroidogenesis.
    • These findings support a cyclic nucleotide-mediated mechanism for ACTH-regulated steroidogenesis.

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