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  • Original Paper
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A conditionally-active form of MEK1 results in autocrine transformation of human and mouse hematopoietic cells

Oncogene volume 19, pages 526–536 (2000)Cite this article

Abstract

The Raf/MEK/MAP kinase cascade plays a critical role in transducing growth signals from activated cell surface receptors. Using ΔMEK1:ER, a conditionally-active form of MEK1, we demonstrate the ability of this dual specificity protein kinase to abrogate the cytokine-dependency of the human and murine hematopoietic cells lines TF-1, FDC-P1 and FL5.12. Cytokine-independent cells were obtained from TF-1, FDC-P1 and FL5.12 cells at frequencies of 2.5×10−3, 5×10−5 and 10−7 respectively, indicating that not all cells expressing ΔMEK1:ER were factor-independent. In general, cells that were converted to a cytokine-independent phenotype displayed a higher level of MAP kinase activity in response to ΔMEK1:ER activation than those that remained cytokine-dependent. ΔMEK1:ER-responsive cells could be maintained long-term in the presence of β-estradiol as well as the estrogen-receptor antagonist 4-Hydroxy-Tamoxifen and the anti-estrogen ICI 164 383. Removal of hormone led to the rapid cessation of cell growth in a manner similar to that observed when cytokine is withdrawn from the parental cells. Treatment of ΔMEK1:ER-responsive cells with a specific and selective inhibitor, PD98059, prevented growth in response to β-estradiol. GM-CSF mRNA transcripts were detected in the MEK1-responsive cells indicating that the activated ΔMEK1:ER may induce a pathway leading to autocrine proliferation. Treatment of MEK1-responsive cells with an anti-GM-CSF antibody, but not a control antibody, suppressed cell growth. The cell lines described here will be useful for elaborating the ability of the MAP kinase pathway to regulate cell proliferation in hematopoietic cells.

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Acknowledgements

We appreciate the art work done by Ms Catherine Spruill and the staff of the ECU Center for Medical Communications. This work was supported in part by grants from the NCI (R01CA51025), the North Carolina Biotechnology Center (9805-ARG-0006) and the East Carolina University School of Medicine Research Grants to (JA McCubrey).

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Author notes

  1. Sean A McCarthy

    Present address: Millennium Pharmaceuticals, Inc., Cambridge, 02139, Massachusetts, MA, USA

  2. Martin McMahon

    Present address: UCSF/Mt. Zion Cancer Center, Box 0128, 2340 Sutter Street, S-329, San Francisco, 94115, California, CA, USA

Authors and Affiliations

  1. Department of Microbiology and Immunology, East Carolina University School of Medicine, Greenville, 27858, North Carolina, NC, USA

    William L Blalock, Marianne Pearce, Linda S Steelman, Richard A Franklin & James A McCubrey

  2. Leo Jenkins Cancer Center, East Carolina University School of Medicine, Greenville, 27858, North Carolina, NC, USA

    Richard A Franklin & James A McCubrey

  3. Department of Cell Signaling, DNAX Research Institute of Molecular and Cellular Biology, 901 California Avenue, Palo Alto, 94304, California, CA, USA

    Sean A McCarthy, Holly Cherwinski & Martin McMahon

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Blalock, W., Pearce, M., Steelman, L. et al. A conditionally-active form of MEK1 results in autocrine transformation of human and mouse hematopoietic cells. Oncogene 19, 526–536 (2000). https://doi.org/10.1038/sj.onc.1203337

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