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  • Molecular Targets For Therapy (MTT)
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Molecular Targets for Therapy (MTT)

Differential effects of kinase cascade inhibitors on neoplastic and cytokine-mediated cell proliferation

Leukemia volume 17, pages 1765–1782 (2003)Cite this article

Abstract

The Raf/MEK/ERK and PI3K/Akt pathways regulate proliferation and prevent apoptosis, and their altered expression is commonly observed in human cancer due to the high mutation frequency of upstream regulators. In this study, the effects of Raf, MEK, and PI3K inhibitors on conditionally transformed hematopoietic cells were examined to determine if they would display cytotoxic differences between cytokine- and oncogene-mediated proliferation, and whether inhibition of both pathways was a more effective means to induce apoptosis. In the hematopoietic model system employed, proliferation was conditional and occurred when either interleukin-3 (IL-3) or the estrogen receptor antagonist 4-hydroxytamoxifen (4HT), which activates the conditional oncoprotein (ΔRaf:ER), were provided. Thus, upon the addition of the signal transduction inhibitors and either IL-3 or 4HT, the effects of these drugs were examined in the same cell under ‘cytokine-’ and ‘oncoprotein’ -mediated growth conditions avoiding genetic and differentiation stage heterogeneity. At drug concentrations around the reported IC50 for the Raf inhibitor L-779,450, it suppressed DNA synthesis and induced apoptosis in hematopoietic FDC-P1 cells transformed to grow in response to either Raf-1 or A-Raf (FD/ΔRaf-1:ER and FD/ΔA-Raf:ER), but it displayed less effects on DNA synthesis and apoptosis when the cells were cultured in IL-3. This Raf inhibitor was less effective on B-Raf- or MEK1-responsive cells, demonstrating the specificity of this drug. MEK inhibitors also suppressed DNA synthesis and induced apoptosis in Raf-responsive cells and the effects were more significant on Raf-responsive compared to cytokine-mediated growth. The PI3K inhibitor LY294002 suppressed Raf-mediated growth, indicating that part of the long-term proliferative effects mediated by Raf are PI3K dependent. Simultaneous inhibition of both Raf/MEK/ERK and PI3K/Akt pathways proved a more efficient means to suppress DNA synthesis and induce apoptosis at lower drug concentrations.

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Acknowledgements

We thank Ms Catherine Spruill for the outstanding artwork. This work was supported in part by a grant form the United States National Institutes of Health (National Cancer Institute, CA51025). We appreciate the comments on this manuscript provided by Dr Fred Bertrand. We thank Merck for generously providing L-779,450.

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  1. J G Shelton and P W Moye: The first two authors contributed equally to this manuscript

Authors and Affiliations

  1. Department of Microbiology and Immunology, Brody School of Medicine, East Carolina University, Greenville, USA

    J G Shelton, P W Moye, L S Steelman, W L Blalock, J T Lee, R A Franklin & J A McCubrey

  2. Leo Jenkins Cancer Center, Brody School of Medicine, East Carolina University, Greenville, USA

    R A Franklin & J A McCubrey

  3. UCSF/Mt. Zion Cancer Center, San Francisco, USA

    M McMahon

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Shelton, J., Moye, P., Steelman, L. et al. Differential effects of kinase cascade inhibitors on neoplastic and cytokine-mediated cell proliferation. Leukemia 17, 1765–1782 (2003). https://doi.org/10.1038/sj.leu.2403052

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