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Signal Transduction

Restoration of SHIP activity in a human leukemia cell line downregulates constitutively activated phosphatidylinositol 3-kinase/Akt/GSK-3β signaling and leads to an increased transit time through the G1 phase of the cell cycle

Leukemia volume 18, pages 1839–1849 (2004)Cite this article

Abstract

The inositol 5-phosphatase SHIP (SHIP-1) is a negative regulator of signal transduction in hematopoietic cells and targeted disruption of SHIP in mice leads to a myeloproliferative disorder. We analyzed the effects of SHIP on the human leukemia cell line Jurkat in which expression of endogenous SHIP protein is not detectable. Restoration of SHIP expression in Jurkat cells with an inducible expression system caused a 69% reduction of phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P3) and a 65% reduction of Akt kinase activity, which was associated with reduced phosphorylation of glycogen synthase kinase 3β (GSK-3β) (Ser-9) without changing the phosphorylation of Bad (Ser-136), FKHR (Ser-256) or MAPK (Thr-202/Tyr-204). SHIP-expressing Jurkat cells showed an increased transit time through the G1 phase of the cell cycle, but SHIP did not cause a complete cell cycle arrest or apoptosis. Extension of the G1 phase was associated with an increased stability of the cell cycle inhibitor p27Kip1 and reduced phosphorylation of the retinoblastoma protein Rb at serine residue 780. Our data indicate that restoration of SHIP activity in a human leukemia cell line, which has lost expression of endogenous SHIP, downregulates constitutively activated phosphatidylinositol 3-kinase/Akt/GSK-3β signaling and leads to an increased transit time through the G1 phase of the cell cycle.

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Acknowledgements

We thank W Wegner, M Engel and B Serreck for excellent technical assistance and C Stocking for her help in constructing the retroviral vector with a tetracycline response element. We also thank H Halfter for supplying antibodies directed against Kip1, C Müller for anti-cyclin antibodies, A Guse for providing the Jurkat Tet-On cells, G Nolan for the Phoenix-Ampho packaging cell line and D von Laer for providing a plasmid encoding the retroviral gag and pol proteins (pSVgp) and the G protein of the vesicular stomatitis virus (pHCMV-VSV-G). This work was supported by grants from the Deutsche Forschungsgemeinschaft to MJ and GWM (JU255/2-3 and JU255/2-4).

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Authors and Affiliations

  1. Center of Experimental Medicine, Institute of Biochemistry and Molecular Biology I: Cellular Signal Transduction, University Hospital Hamburg-Eppendorf, Hamburg, Germany

    S Horn, M M Weck, G W Mayr & M Jücker

  2. Department of Immunology and Cell Biology, Research Center Borstel, Borstel, Germany

    E Endl

  3. Bone Marrow Transplantation Center, University Hospital Hamburg-Eppendorf, Hamburg, Germany

    B Fehse

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  1. S Horn
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Correspondence to M Jücker.

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Horn, S., Endl, E., Fehse, B. et al. Restoration of SHIP activity in a human leukemia cell line downregulates constitutively activated phosphatidylinositol 3-kinase/Akt/GSK-3β signaling and leads to an increased transit time through the G1 phase of the cell cycle. Leukemia 18, 1839–1849 (2004). https://doi.org/10.1038/sj.leu.2403529

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