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Hypoosmotic stress stimulates growth in HepG2 cells via protein kinase B-dependent activation of activator protein-1

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Journal of Gastrointestinal Surgery

Abstract

Although hypoosmotic stress-induced cell swelling activates phosphatidylinositol-3-kinase, its impact on the downstream signal protein kinase B and cell growth is unknown. Activator protein-1 is in part phosphatidylinositol-3-kinase dependent, and is important in proliferation. We hypothesized that cell swelling modulates proliferation in HepG2 cells via the protein kinase B-dependent activation of activator protein-1. HepG2 cells pretreated with or without LY294002 were exposed for up to 30 minutes to hypoosmotic medium (160 mOsm/L). Tumor necrosis factor-alpha (1.4 nmol/L) or normoosmolar medium (270 mOsm/L) served as positive and negative controls, respectively. Western immunoblots measured cytoplasmic phosphorylated and total protein kinase B. Electromobility shift assays measured nuclear activator protein-1. Methylene blue assays measured cell proliferation at 24, 48, and 72 hours after stimulation. Hypoosmotic stress phosphorylated protein kinase B by 10 minutes. Subsequently, hypoosmotic exposure stimulated activator protein-1 by 30 minutes. Pulse exposure to hypoosmotic stress potentiated HepG2 proliferation by 72 hours as compared to both negative controls and LY-inhibited cells (n = 4 per group, P = 0.009 and P = 0.004, respectively; P <0.001 analysis of variance. All three activation events were abolished with LY294002 pretreatment. In HepG2 cells, hypoosmotic stress-induced swelling stimulates proliferation via protein kinase B-mediated activation of activator protein-1. These data delineate a possible mechanism linking changes in cell volume to growth in human liver cancer.

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Authors and Affiliations

  1. Departments of Surgery, University of Massa-chusetts Medical School, 55 Lake Avenue North, 01655, Worcester, MA

    Robin D. Kim M.D., Timothy P. Roth M.S., Chad E. Darling M.D., Rocco Ricciardi M.D., Bradley K. Schaffer M.D. & Ravi S. Chart M.D. (Assistant Professor of Surgery and Cell Biology)

  2. Departments of Cell Biology, University of Massa-chusetts Medical School, Worcester, Mass

    Ravi S. Chart M.D. (Assistant Professor of Surgery and Cell Biology)

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  1. Robin D. Kim M.D.
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  2. Timothy P. Roth M.S.
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  3. Chad E. Darling M.D.
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  4. Rocco Ricciardi M.D.
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  5. Bradley K. Schaffer M.D.
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Correspondence to Ravi S. Chart M.D..

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Kim, R.D., Roth, T.P., Darling, C.E. et al. Hypoosmotic stress stimulates growth in HepG2 cells via protein kinase B-dependent activation of activator protein-1. J Gastrointest Surg 5, 546–555 (2001). https://doi.org/10.1016/S1091-255X(01)80094-9

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  • DOI: https://doi.org/10.1016/S1091-255X(01)80094-9

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