C/EBPα inactivation in FAK-overexpressed HL-60 cells impairs cell differentiation

Ken Ichiro Hashimoto, Yoshiko Sonoda, Masakazu Yamakado, Megumi Tago, Naomi Yoshida, Akiko Rokudai, Eriko Yokota, Tadashi Kasahara

Research output: Contribution to journalArticle

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Abstract

We previously demonstrated that focal adhesion kinase (FAK)-overexpressed (HL-60/FAK) cells have marked resistance against various apoptotic stimuli such as oxidative stress, ionizing radiation and TNF-receptor-induced ligand (TRAIL) compared with vector-transfected (HL-60/Vect) cells. Here, we show that HL-60/FAK cells are highly resistant to all-trans retinoic acid (ATRA)-induced differentiation, whereas original HL-60 or HL-60/Vect cells are sensitive. Treatment with ATRA at 1 μM for 5 days markedly inhibited the proliferation and increased the expression of differentiation markers (CD38, CD11b) in HL-60/Vect cells, but showed no such effect in HL-60/FAK cells. Electrophoretic mobility shift assay (EMSA) using an oligonucleotide for the c/EBP consensus binding sequence showed that c/EBPα was activated in ATRA-treated HL-60/Vect cells but not in HL-60/FAK cells, indicating that c/EBPα activation by ATRA was impaired in HL-60/FAK cells. In addition, the association of retinoblastoma protein (pRb) and c/EBPα after treatment with ATRA was seen in HL-60/Vect cells but not in HL-60/FAK cells. Further, hyperphosphorylation of pRb was observed in HL-60/FAK cells. Finally, the introduction of FAK siRNA into HL-60/FAK cells resulted in the recovery of sensitivity to ATRA-induced differentiation, confirming that the inhibition of HL-60/FAK differentiation resulted from both the induction of pRb hyperphosphorylation and the inhibition of association of pRb and c/EBPα.

Original languageEnglish
Pages (from-to)955-963
Number of pages9
JournalCellular Signalling
Volume18
Issue number7
DOIs
Publication statusPublished - 2006 Jul

Fingerprint

Focal Adhesion Protein-Tyrosine Kinases
HL-60 Cells
Cell Differentiation
Tretinoin
Tumor Necrosis Factors
Retinoblastoma Protein
Differentiation Antigens
Consensus Sequence
Electrophoretic Mobility Shift Assay
Ionizing Radiation
Oligonucleotides
Small Interfering RNA
Oxidative Stress

Keywords

  • c/EBP
  • Differentiation
  • FAK
  • Granulocyte
  • pRb

ASJC Scopus subject areas

  • Cell Biology

Cite this

Hashimoto, K. I., Sonoda, Y., Yamakado, M., Tago, M., Yoshida, N., Rokudai, A., ... Kasahara, T. (2006). C/EBPα inactivation in FAK-overexpressed HL-60 cells impairs cell differentiation. Cellular Signalling, 18(7), 955-963. https://doi.org/10.1016/j.cellsig.2005.08.014

C/EBPα inactivation in FAK-overexpressed HL-60 cells impairs cell differentiation. / Hashimoto, Ken Ichiro; Sonoda, Yoshiko; Yamakado, Masakazu; Tago, Megumi; Yoshida, Naomi; Rokudai, Akiko; Yokota, Eriko; Kasahara, Tadashi.

In: Cellular Signalling, Vol. 18, No. 7, 07.2006, p. 955-963.

Research output: Contribution to journalArticle

Hashimoto, KI, Sonoda, Y, Yamakado, M, Tago, M, Yoshida, N, Rokudai, A, Yokota, E & Kasahara, T 2006, 'C/EBPα inactivation in FAK-overexpressed HL-60 cells impairs cell differentiation', Cellular Signalling, vol. 18, no. 7, pp. 955-963. https://doi.org/10.1016/j.cellsig.2005.08.014
Hashimoto, Ken Ichiro ; Sonoda, Yoshiko ; Yamakado, Masakazu ; Tago, Megumi ; Yoshida, Naomi ; Rokudai, Akiko ; Yokota, Eriko ; Kasahara, Tadashi. / C/EBPα inactivation in FAK-overexpressed HL-60 cells impairs cell differentiation. In: Cellular Signalling. 2006 ; Vol. 18, No. 7. pp. 955-963.
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AU - Sonoda, Yoshiko

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AU - Tago, Megumi

AU - Yoshida, Naomi

AU - Rokudai, Akiko

AU - Yokota, Eriko

AU - Kasahara, Tadashi

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AB - We previously demonstrated that focal adhesion kinase (FAK)-overexpressed (HL-60/FAK) cells have marked resistance against various apoptotic stimuli such as oxidative stress, ionizing radiation and TNF-receptor-induced ligand (TRAIL) compared with vector-transfected (HL-60/Vect) cells. Here, we show that HL-60/FAK cells are highly resistant to all-trans retinoic acid (ATRA)-induced differentiation, whereas original HL-60 or HL-60/Vect cells are sensitive. Treatment with ATRA at 1 μM for 5 days markedly inhibited the proliferation and increased the expression of differentiation markers (CD38, CD11b) in HL-60/Vect cells, but showed no such effect in HL-60/FAK cells. Electrophoretic mobility shift assay (EMSA) using an oligonucleotide for the c/EBP consensus binding sequence showed that c/EBPα was activated in ATRA-treated HL-60/Vect cells but not in HL-60/FAK cells, indicating that c/EBPα activation by ATRA was impaired in HL-60/FAK cells. In addition, the association of retinoblastoma protein (pRb) and c/EBPα after treatment with ATRA was seen in HL-60/Vect cells but not in HL-60/FAK cells. Further, hyperphosphorylation of pRb was observed in HL-60/FAK cells. Finally, the introduction of FAK siRNA into HL-60/FAK cells resulted in the recovery of sensitivity to ATRA-induced differentiation, confirming that the inhibition of HL-60/FAK differentiation resulted from both the induction of pRb hyperphosphorylation and the inhibition of association of pRb and c/EBPα.

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