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, M. Suzuki, M. Nakamura, Y. Banno, Y. Nozawa, T. Murate

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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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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, 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, 1833(5), 1006-1016. https://doi.org/10.1016/j.bbamcr.2013.01.001