BimEL is an important determinant for induction of anoikis sensitivity by mitogen-activated protein/extracellular signal-regulated kinase kinase inhibitors

Hidesuke Fukazawa, Kohji Noguchi, Atsuko Masumi, Yuko Murakami, Yoshimasa Uehara

Research output: Contribution to journalArticle

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Abstract

Loss of contact with substratum triggers apoptosis in many normal cell types, a phenomenon termed anoikis. We reported previously that mitogen-activated protein/ extracellular signal-regulated kinase kinase (MEK) inhibitors induced apoptosis in nonanchored MDA-MB231 and HBC4 human breast cancer cells, whereas anchored cells remained viable. Here, we report that activation of the BH3-only protein BimEL is the major mechanism for induction of anoikis sensitivity by MEK inhibitors in MDA-MB231 and HBC4 cells. On treatment with MEK inhibitors, BimEL in MDA-MB231 and HBC4 cells rapidly increased, irrespective of the state of anchorage. However, it translocated to mitochondria only in nonanchored cells, explaining why attached cells remain viable. MDAMB231 and HBC4 cells had exceedingly low basal levels of BimEL compared with other breast cancer cells, suggesting that maintenance of low BimEL amount is important for survival of these cells. MEK inhibitors also induced the electrophoretic mobility shift of BimEL, indicative of reduced phosphorylation. In vitro, BimEL was phosphorylated by extracellular signal-regulated kinase on Ser69, which resides in the BimEL-specific insert region. Using phosphospecific antibody against this site, we show that this residue is actually phosphorylated in cells. We also show that phosphorylation of Ser69 promotes ubiquitination of BimEL. We conclude that MEK inhibitors sensitize MDA-MB231 and HBC4 cells to anoikis by blocking phosphorylation and hence degradation of BimEL, a mechanism that these cells depend on to escape anoikis.

Original languageEnglish
Pages (from-to)1281-1288
Number of pages8
JournalMolecular Cancer Therapeutics
Volume3
Issue number10
Publication statusPublished - 2004 Oct
Externally publishedYes

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Anoikis
Extracellular Signal-Regulated MAP Kinases
Mitogens
Phosphotransferases
Proteins
Mitogen-Activated Protein Kinase Kinases
Phosphorylation
Phospho-Specific Antibodies
Apoptosis
Breast Neoplasms
MAP Kinase Kinase Kinases
Ubiquitination

ASJC Scopus subject areas

  • Oncology
  • Drug Discovery
  • Pharmacology

Cite this

BimEL is an important determinant for induction of anoikis sensitivity by mitogen-activated protein/extracellular signal-regulated kinase kinase inhibitors. / Fukazawa, Hidesuke; Noguchi, Kohji; Masumi, Atsuko; Murakami, Yuko; Uehara, Yoshimasa.

In: Molecular Cancer Therapeutics, Vol. 3, No. 10, 10.2004, p. 1281-1288.

Research output: Contribution to journalArticle

Fukazawa, Hidesuke ; Noguchi, Kohji ; Masumi, Atsuko ; Murakami, Yuko ; Uehara, Yoshimasa. / BimEL is an important determinant for induction of anoikis sensitivity by mitogen-activated protein/extracellular signal-regulated kinase kinase inhibitors. In: Molecular Cancer Therapeutics. 2004 ; Vol. 3, No. 10. pp. 1281-1288.
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