Sphingosine kinase 1 expression is downregulated during differentiation of friend cells due to decreased c-MYB

N. Mizutani, M. Kobayashi, S. Sobue, M. Ichihara, H. Ito, K. Tanaka, S. Iwaki, S. Fujii, Y. Ito, K. Tamiya-Koizumi, A. Takagi, T. Kojima, T. Naoe, Motoshi Suzuki, M. Nakamura, Y. Banno, Y. Nozawa, T. Murate

Research output: Contribution to journalArticle

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Abstract

Sphingosine kinase 1 (SPHK1) overexpression in malignant cells has been reported. Mouse Friend cells showed higher SPHK1 but not SPHK2 expression compared with other mouse cell lines. A Sphk1 promoter analysis demonstrated the region between -. 53. bp and the first exon as the minimal promoter. Further promoter truncation revealed the importance of a MYB-binding site. EMSA using this region as the probe demonstrated one band containing c-MYB protein, and its intensity decreased during erythroid differentiation with hexamethylane bisacetamide (HMBA), a potent inducer of erythroid differentiation of Friend cells. ChIP assay also revealed in vivo binding of c-MYB. c-MYB overexpression and siRNA for c-Myb affected SPHK1 expression, confirming the important regulatory role of c-MYB in SPHK1 expression. HMBA reduced c-MYB expression rapidly. Induced differentiation by HMBA caused a marked and rapid reduction of SPHK1 mRNA, protein and enzyme activity leading to the rapid decrease of cellular sphingosine 1-phosphate level. Moreover, terminally differentiated cells did not resume SPHK1 expression. Compared with original Friend cells, stable overexpression of wild-type SPHK1 showed higher cell proliferation, resistance to cell death by serum depletion. Interestingly, HMBA-induced differentiation of these cells was delayed but not completely suppressed. In contrast, SPHK inhibitor and its siRNA inhibited cell growth and enhanced HMBA-induced differentiation significantly, suggesting that SPHK1 delayed HMBA-induced differentiation by its cell proliferation-promoting activity. Effects of pertussis toxin, a G-protein-coupled receptor inhibitor, and S1P receptor antagonist on Friend cell growth and differentiation were negligible, suggesting the importance of the intracellular SPHK1/S1P signaling in Friend cells.

Original languageEnglish
Pages (from-to)1006-1016
Number of pages11
JournalBiochimica et Biophysica Acta - Molecular Cell Research
Volume1833
Issue number5
DOIs
Publication statusPublished - 01-05-2013
Externally publishedYes

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Cell Differentiation
Down-Regulation
Small Interfering RNA
Cell Proliferation
Lysosphingolipid Receptors
sphingosine kinase
Pertussis Toxin
Growth
G-Protein-Coupled Receptors
Exons
Proteins
Cell Death
Binding Sites
Cell Line
Messenger RNA
Enzymes
Serum

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

Cite this

Mizutani, N. ; Kobayashi, M. ; Sobue, S. ; Ichihara, M. ; Ito, H. ; Tanaka, K. ; Iwaki, S. ; Fujii, S. ; Ito, Y. ; Tamiya-Koizumi, K. ; Takagi, A. ; Kojima, T. ; Naoe, T. ; Suzuki, Motoshi ; Nakamura, M. ; Banno, Y. ; Nozawa, Y. ; Murate, T. / Sphingosine kinase 1 expression is downregulated during differentiation of friend cells due to decreased c-MYB. In: Biochimica et Biophysica Acta - Molecular Cell Research. 2013 ; Vol. 1833, No. 5. pp. 1006-1016.
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title = "Sphingosine kinase 1 expression is downregulated during differentiation of friend cells due to decreased c-MYB",
abstract = "Sphingosine kinase 1 (SPHK1) overexpression in malignant cells has been reported. Mouse Friend cells showed higher SPHK1 but not SPHK2 expression compared with other mouse cell lines. A Sphk1 promoter analysis demonstrated the region between -. 53. bp and the first exon as the minimal promoter. Further promoter truncation revealed the importance of a MYB-binding site. EMSA using this region as the probe demonstrated one band containing c-MYB protein, and its intensity decreased during erythroid differentiation with hexamethylane bisacetamide (HMBA), a potent inducer of erythroid differentiation of Friend cells. ChIP assay also revealed in vivo binding of c-MYB. c-MYB overexpression and siRNA for c-Myb affected SPHK1 expression, confirming the important regulatory role of c-MYB in SPHK1 expression. HMBA reduced c-MYB expression rapidly. Induced differentiation by HMBA caused a marked and rapid reduction of SPHK1 mRNA, protein and enzyme activity leading to the rapid decrease of cellular sphingosine 1-phosphate level. Moreover, terminally differentiated cells did not resume SPHK1 expression. Compared with original Friend cells, stable overexpression of wild-type SPHK1 showed higher cell proliferation, resistance to cell death by serum depletion. Interestingly, HMBA-induced differentiation of these cells was delayed but not completely suppressed. In contrast, SPHK inhibitor and its siRNA inhibited cell growth and enhanced HMBA-induced differentiation significantly, suggesting that SPHK1 delayed HMBA-induced differentiation by its cell proliferation-promoting activity. Effects of pertussis toxin, a G-protein-coupled receptor inhibitor, and S1P receptor antagonist on Friend cell growth and differentiation were negligible, suggesting the importance of the intracellular SPHK1/S1P signaling in Friend cells.",
author = "N. Mizutani and M. Kobayashi and S. Sobue and M. Ichihara and H. Ito and K. Tanaka and S. Iwaki and S. Fujii and Y. Ito and K. Tamiya-Koizumi and A. Takagi and T. Kojima and T. Naoe and Motoshi Suzuki and M. Nakamura and Y. Banno and Y. Nozawa and T. Murate",
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Mizutani, N, Kobayashi, M, Sobue, S, Ichihara, M, Ito, H, Tanaka, K, Iwaki, S, Fujii, S, Ito, Y, Tamiya-Koizumi, K, Takagi, A, Kojima, T, Naoe, T, Suzuki, M, Nakamura, M, Banno, Y, Nozawa, Y & Murate, T 2013, 'Sphingosine kinase 1 expression is downregulated during differentiation of friend cells due to decreased c-MYB', Biochimica et Biophysica Acta - Molecular Cell Research, vol. 1833, no. 5, pp. 1006-1016. https://doi.org/10.1016/j.bbamcr.2013.01.001

Sphingosine kinase 1 expression is downregulated during differentiation of friend cells due to decreased c-MYB. / Mizutani, N.; Kobayashi, M.; Sobue, S.; Ichihara, M.; Ito, H.; Tanaka, K.; Iwaki, S.; Fujii, S.; Ito, Y.; Tamiya-Koizumi, K.; Takagi, A.; Kojima, T.; Naoe, T.; Suzuki, Motoshi; Nakamura, M.; Banno, Y.; Nozawa, Y.; Murate, T.

In: Biochimica et Biophysica Acta - Molecular Cell Research, Vol. 1833, No. 5, 01.05.2013, p. 1006-1016.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Sphingosine kinase 1 expression is downregulated during differentiation of friend cells due to decreased c-MYB

AU - Mizutani, N.

AU - Kobayashi, M.

AU - Sobue, S.

AU - Ichihara, M.

AU - Ito, H.

AU - Tanaka, K.

AU - Iwaki, S.

AU - Fujii, S.

AU - Ito, Y.

AU - Tamiya-Koizumi, K.

AU - Takagi, A.

AU - Kojima, T.

AU - Naoe, T.

AU - Suzuki, Motoshi

AU - Nakamura, M.

AU - Banno, Y.

AU - Nozawa, Y.

AU - Murate, T.

PY - 2013/5/1

Y1 - 2013/5/1

N2 - Sphingosine kinase 1 (SPHK1) overexpression in malignant cells has been reported. Mouse Friend cells showed higher SPHK1 but not SPHK2 expression compared with other mouse cell lines. A Sphk1 promoter analysis demonstrated the region between -. 53. bp and the first exon as the minimal promoter. Further promoter truncation revealed the importance of a MYB-binding site. EMSA using this region as the probe demonstrated one band containing c-MYB protein, and its intensity decreased during erythroid differentiation with hexamethylane bisacetamide (HMBA), a potent inducer of erythroid differentiation of Friend cells. ChIP assay also revealed in vivo binding of c-MYB. c-MYB overexpression and siRNA for c-Myb affected SPHK1 expression, confirming the important regulatory role of c-MYB in SPHK1 expression. HMBA reduced c-MYB expression rapidly. Induced differentiation by HMBA caused a marked and rapid reduction of SPHK1 mRNA, protein and enzyme activity leading to the rapid decrease of cellular sphingosine 1-phosphate level. Moreover, terminally differentiated cells did not resume SPHK1 expression. Compared with original Friend cells, stable overexpression of wild-type SPHK1 showed higher cell proliferation, resistance to cell death by serum depletion. Interestingly, HMBA-induced differentiation of these cells was delayed but not completely suppressed. In contrast, SPHK inhibitor and its siRNA inhibited cell growth and enhanced HMBA-induced differentiation significantly, suggesting that SPHK1 delayed HMBA-induced differentiation by its cell proliferation-promoting activity. Effects of pertussis toxin, a G-protein-coupled receptor inhibitor, and S1P receptor antagonist on Friend cell growth and differentiation were negligible, suggesting the importance of the intracellular SPHK1/S1P signaling in Friend cells.

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