Effects of the RAF/MEK/ERK and PI3k/AKT signal transduction pathways on the abrogation of cytokine-dependence and prevention of apoptosis in hematopoietic cells

John G. Shelton, Linda S. Steelman, John T. Lee, Steven L. Knapp, William L. Blalock, Phillip W. Moye, Richard A. Franklin, Steven C. Pohnert, Amer M. Mirza, Martin McMahon, James A. McCubrey

Research output: Contribution to journalArticle

97 Citations (Scopus)

Abstract

The Raf/MEK/ERK kinase cascade is pivotal in transmitting signals from membrane receptors to transcription factors that control gene expression culminating in the regulation of cell cycle progression. This cascade can prevent cell death through ERK2 and p90Rsk activation and phosphorylation of apoptotic and cell cycle regulatory proteins. The PI3K/Akt kinase cascade also controls apoptosis and can phosphorylate many apoptotic and cell cycle regulatory proteins. These pathways are interwoven as Akt can phosphorylate Raf and result in its inactivation, and Raf can be required for the antiapoptotic effects of Akt. In this study, the effects of activated Raf (Raf-1, A-Raf and B-Raf) and PI3K/Akt proteins on abrogation of cytokine dependence in FL5.12 hematopoietic cells were examined. Activated Raf, PI3K or Akt expression, by themselves, did not readily relieve cytokine dependence. The presence of activated Raf and PI3K/Akt increased the isolation of factor-independent cells from 400- to 2500-fold depending upon the particular combination examined. The individual effects of activated Raf and Akt on proliferation, apoptosis and autocrine growth factor synthesis were further examined with hormone-inducible constructs (ΔRaf-I:AR and ΔAkt:ER*(Myr+). Activation of either Raf or Akt hindered cell death; however, both proliferation and maximal synthesis of autocrine cytokines were dependent upon activation of both signaling pathways. The effects of small molecular weight inhibitors on DNA synthesis and cytokine gene expression were also examined. The PI3K inhibitor, LY294002, inhibited growth and cytokine gene expression. This effect could be synergistically increased by addition of the MEK inhibitor UO126. These cells will be useful in elucidating the interactions between Raf/MEK/ERK and PI3K/Akt cascades in proliferation, apoptosis, and leukemogenesis, as well as evaluating the efficacy of signal transduction inhibitors that target these cascades.

Original languageEnglish
Pages (from-to)2478-2492
Number of pages15
JournalOncogene
Volume22
Issue number16
DOIs
Publication statusPublished - Apr 24 2003

Fingerprint

Mitogen-Activated Protein Kinase Kinases
Phosphatidylinositol 3-Kinases
Signal Transduction
Apoptosis
Cytokines
Cell Cycle Proteins
Gene Expression
Cell Death
Nucleic Acid Synthesis Inhibitors
MAP Kinase Kinase Kinases
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Intercellular Signaling Peptides and Proteins
Cell Cycle
Transcription Factors
Phosphotransferases
Molecular Weight
Phosphorylation
Hormones
Membranes
Growth

Keywords

  • Akt
  • Annexin V binding
  • Apoptosis
  • Autocrine transformation
  • Cytokine dependence
  • Hematopoietic cells
  • P13K
  • Raf
  • Signal transduction

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research
  • Genetics

Cite this

Effects of the RAF/MEK/ERK and PI3k/AKT signal transduction pathways on the abrogation of cytokine-dependence and prevention of apoptosis in hematopoietic cells. / Shelton, John G.; Steelman, Linda S.; Lee, John T.; Knapp, Steven L.; Blalock, William L.; Moye, Phillip W.; Franklin, Richard A.; Pohnert, Steven C.; Mirza, Amer M.; McMahon, Martin; McCubrey, James A.

In: Oncogene, Vol. 22, No. 16, 24.04.2003, p. 2478-2492.

Research output: Contribution to journalArticle

Shelton, JG, Steelman, LS, Lee, JT, Knapp, SL, Blalock, WL, Moye, PW, Franklin, RA, Pohnert, SC, Mirza, AM, McMahon, M & McCubrey, JA 2003, 'Effects of the RAF/MEK/ERK and PI3k/AKT signal transduction pathways on the abrogation of cytokine-dependence and prevention of apoptosis in hematopoietic cells', Oncogene, vol. 22, no. 16, pp. 2478-2492. https://doi.org/10.1038/sj.onc.1206321
Shelton, John G. ; Steelman, Linda S. ; Lee, John T. ; Knapp, Steven L. ; Blalock, William L. ; Moye, Phillip W. ; Franklin, Richard A. ; Pohnert, Steven C. ; Mirza, Amer M. ; McMahon, Martin ; McCubrey, James A. / Effects of the RAF/MEK/ERK and PI3k/AKT signal transduction pathways on the abrogation of cytokine-dependence and prevention of apoptosis in hematopoietic cells. In: Oncogene. 2003 ; Vol. 22, No. 16. pp. 2478-2492.
@article{6d162394c63f41c6907d19bcca23bece,
title = "Effects of the RAF/MEK/ERK and PI3k/AKT signal transduction pathways on the abrogation of cytokine-dependence and prevention of apoptosis in hematopoietic cells",
abstract = "The Raf/MEK/ERK kinase cascade is pivotal in transmitting signals from membrane receptors to transcription factors that control gene expression culminating in the regulation of cell cycle progression. This cascade can prevent cell death through ERK2 and p90Rsk activation and phosphorylation of apoptotic and cell cycle regulatory proteins. The PI3K/Akt kinase cascade also controls apoptosis and can phosphorylate many apoptotic and cell cycle regulatory proteins. These pathways are interwoven as Akt can phosphorylate Raf and result in its inactivation, and Raf can be required for the antiapoptotic effects of Akt. In this study, the effects of activated Raf (Raf-1, A-Raf and B-Raf) and PI3K/Akt proteins on abrogation of cytokine dependence in FL5.12 hematopoietic cells were examined. Activated Raf, PI3K or Akt expression, by themselves, did not readily relieve cytokine dependence. The presence of activated Raf and PI3K/Akt increased the isolation of factor-independent cells from 400- to 2500-fold depending upon the particular combination examined. The individual effects of activated Raf and Akt on proliferation, apoptosis and autocrine growth factor synthesis were further examined with hormone-inducible constructs (ΔRaf-I:AR and ΔAkt:ER*(Myr+). Activation of either Raf or Akt hindered cell death; however, both proliferation and maximal synthesis of autocrine cytokines were dependent upon activation of both signaling pathways. The effects of small molecular weight inhibitors on DNA synthesis and cytokine gene expression were also examined. The PI3K inhibitor, LY294002, inhibited growth and cytokine gene expression. This effect could be synergistically increased by addition of the MEK inhibitor UO126. These cells will be useful in elucidating the interactions between Raf/MEK/ERK and PI3K/Akt cascades in proliferation, apoptosis, and leukemogenesis, as well as evaluating the efficacy of signal transduction inhibitors that target these cascades.",
keywords = "Akt, Annexin V binding, Apoptosis, Autocrine transformation, Cytokine dependence, Hematopoietic cells, P13K, Raf, Signal transduction",
author = "Shelton, {John G.} and Steelman, {Linda S.} and Lee, {John T.} and Knapp, {Steven L.} and Blalock, {William L.} and Moye, {Phillip W.} and Franklin, {Richard A.} and Pohnert, {Steven C.} and Mirza, {Amer M.} and Martin McMahon and McCubrey, {James A.}",
year = "2003",
month = "4",
day = "24",
doi = "10.1038/sj.onc.1206321",
language = "English",
volume = "22",
pages = "2478--2492",
journal = "Oncogene",
issn = "0950-9232",
publisher = "Nature Publishing Group",
number = "16",

}

TY - JOUR

T1 - Effects of the RAF/MEK/ERK and PI3k/AKT signal transduction pathways on the abrogation of cytokine-dependence and prevention of apoptosis in hematopoietic cells

AU - Shelton, John G.

AU - Steelman, Linda S.

AU - Lee, John T.

AU - Knapp, Steven L.

AU - Blalock, William L.

AU - Moye, Phillip W.

AU - Franklin, Richard A.

AU - Pohnert, Steven C.

AU - Mirza, Amer M.

AU - McMahon, Martin

AU - McCubrey, James A.

PY - 2003/4/24

Y1 - 2003/4/24

N2 - The Raf/MEK/ERK kinase cascade is pivotal in transmitting signals from membrane receptors to transcription factors that control gene expression culminating in the regulation of cell cycle progression. This cascade can prevent cell death through ERK2 and p90Rsk activation and phosphorylation of apoptotic and cell cycle regulatory proteins. The PI3K/Akt kinase cascade also controls apoptosis and can phosphorylate many apoptotic and cell cycle regulatory proteins. These pathways are interwoven as Akt can phosphorylate Raf and result in its inactivation, and Raf can be required for the antiapoptotic effects of Akt. In this study, the effects of activated Raf (Raf-1, A-Raf and B-Raf) and PI3K/Akt proteins on abrogation of cytokine dependence in FL5.12 hematopoietic cells were examined. Activated Raf, PI3K or Akt expression, by themselves, did not readily relieve cytokine dependence. The presence of activated Raf and PI3K/Akt increased the isolation of factor-independent cells from 400- to 2500-fold depending upon the particular combination examined. The individual effects of activated Raf and Akt on proliferation, apoptosis and autocrine growth factor synthesis were further examined with hormone-inducible constructs (ΔRaf-I:AR and ΔAkt:ER*(Myr+). Activation of either Raf or Akt hindered cell death; however, both proliferation and maximal synthesis of autocrine cytokines were dependent upon activation of both signaling pathways. The effects of small molecular weight inhibitors on DNA synthesis and cytokine gene expression were also examined. The PI3K inhibitor, LY294002, inhibited growth and cytokine gene expression. This effect could be synergistically increased by addition of the MEK inhibitor UO126. These cells will be useful in elucidating the interactions between Raf/MEK/ERK and PI3K/Akt cascades in proliferation, apoptosis, and leukemogenesis, as well as evaluating the efficacy of signal transduction inhibitors that target these cascades.

AB - The Raf/MEK/ERK kinase cascade is pivotal in transmitting signals from membrane receptors to transcription factors that control gene expression culminating in the regulation of cell cycle progression. This cascade can prevent cell death through ERK2 and p90Rsk activation and phosphorylation of apoptotic and cell cycle regulatory proteins. The PI3K/Akt kinase cascade also controls apoptosis and can phosphorylate many apoptotic and cell cycle regulatory proteins. These pathways are interwoven as Akt can phosphorylate Raf and result in its inactivation, and Raf can be required for the antiapoptotic effects of Akt. In this study, the effects of activated Raf (Raf-1, A-Raf and B-Raf) and PI3K/Akt proteins on abrogation of cytokine dependence in FL5.12 hematopoietic cells were examined. Activated Raf, PI3K or Akt expression, by themselves, did not readily relieve cytokine dependence. The presence of activated Raf and PI3K/Akt increased the isolation of factor-independent cells from 400- to 2500-fold depending upon the particular combination examined. The individual effects of activated Raf and Akt on proliferation, apoptosis and autocrine growth factor synthesis were further examined with hormone-inducible constructs (ΔRaf-I:AR and ΔAkt:ER*(Myr+). Activation of either Raf or Akt hindered cell death; however, both proliferation and maximal synthesis of autocrine cytokines were dependent upon activation of both signaling pathways. The effects of small molecular weight inhibitors on DNA synthesis and cytokine gene expression were also examined. The PI3K inhibitor, LY294002, inhibited growth and cytokine gene expression. This effect could be synergistically increased by addition of the MEK inhibitor UO126. These cells will be useful in elucidating the interactions between Raf/MEK/ERK and PI3K/Akt cascades in proliferation, apoptosis, and leukemogenesis, as well as evaluating the efficacy of signal transduction inhibitors that target these cascades.

KW - Akt

KW - Annexin V binding

KW - Apoptosis

KW - Autocrine transformation

KW - Cytokine dependence

KW - Hematopoietic cells

KW - P13K

KW - Raf

KW - Signal transduction

UR - http://www.scopus.com/inward/record.url?scp=0038476605&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0038476605&partnerID=8YFLogxK

U2 - 10.1038/sj.onc.1206321

DO - 10.1038/sj.onc.1206321

M3 - Article

C2 - 12717425

AN - SCOPUS:0038476605

VL - 22

SP - 2478

EP - 2492

JO - Oncogene

JF - Oncogene

SN - 0950-9232

IS - 16

ER -