Role of down-regulated neutral ceramidase during all-trans retinoic acid-induced neuronal differentiation in SH-SY5Y neuroblastoma cells

Kouji Tanaka, Keiko Tamiya-Koizumi, Kazumi Hagiwara, Hiromi Ito, Akira Takagi, Tetsuhito Kojima, Motoshi Suzuki, Soichiro Iwaki, Satoshi Fujii, Mitsuhiro Nakamura, Yoshiko Banno, Reiji Kannagi, Tatsuya Tsurumi, Mamoru Kyogashima, Takashi Murate

Research output: Contribution to journalArticle

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Abstract

Neutral ceramidase (NCDase) is considered to be a critical enzyme for controlling the turnover of ceramide, an important bioactive lipid, which determines cell's fate. All-trans retinoic acid (ATRA) has been reported to induce neuronal differentiation and cell-cycle arrest [Lopez-Carballo, Moreno, Masia, Perez, and Barettino (Activation of the phosphatidylinositol 3-kinase/Akt signalling pathway by retinoic acid is required for neural differentiation of SH-SY5Y human neuroblastoma cells. J Biol Chem 2002:277:25297-304.)]. In this study, we observed that ATRA-induced cellular ceramide accumulation, cell-growth arrest and differentiation accompanied with down-regulation of NCDase in SH-SY5Y cells, without a decrease in sphingosine or sphingosine 1-phosphate. We examined whether the down-regulation of NCDase was involved in the increase in ceramide and cell differentiation. ATRA was found to down-regulate mRNA, protein and the enzyme activity of NCDase. Interestingly, GATA-2 was also decreased with ATRA treatment, and experiments using its expression vector and siRNA and chromatin immunoprecipitation assay demonstrated GATA-2 acted as transcription-factor of NCDase gene expression. By establishing stable transfectants with decreased NCDase expression and activity, we clarified the significance of NCDase down-regulation for ATRA-induced neuronal differentiation. Those sub-clones showed both increased cellular ceramide and reduced cell growth as well as neuronal differentiation phenotypes. These results demonstrate that down-regulation of NCDase through ATRA-induced GATA-2 decrease plays an important role in induction of ceramide accumulation and neuronal differentiation in SH-SY5Y cells.

Original languageEnglish
Pages (from-to)611-620
Number of pages10
JournalJournal of Biochemistry
Volume151
Issue number6
DOIs
Publication statusPublished - 01-06-2012
Externally publishedYes

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Neutral Ceramidase
Tretinoin
Neuroblastoma
Ceramides
Down-Regulation
Cell growth
Phosphatidylinositol 3-Kinase
Sphingosine
Chromatin Immunoprecipitation
Enzyme activity
Enzymes
Growth
Cell Cycle Checkpoints
Gene expression
Small Interfering RNA
Chromatin
Cell Differentiation
Assays
Transcription Factors
Clone Cells

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Tanaka, Kouji ; Tamiya-Koizumi, Keiko ; Hagiwara, Kazumi ; Ito, Hiromi ; Takagi, Akira ; Kojima, Tetsuhito ; Suzuki, Motoshi ; Iwaki, Soichiro ; Fujii, Satoshi ; Nakamura, Mitsuhiro ; Banno, Yoshiko ; Kannagi, Reiji ; Tsurumi, Tatsuya ; Kyogashima, Mamoru ; Murate, Takashi. / Role of down-regulated neutral ceramidase during all-trans retinoic acid-induced neuronal differentiation in SH-SY5Y neuroblastoma cells. In: Journal of Biochemistry. 2012 ; Vol. 151, No. 6. pp. 611-620.
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abstract = "Neutral ceramidase (NCDase) is considered to be a critical enzyme for controlling the turnover of ceramide, an important bioactive lipid, which determines cell's fate. All-trans retinoic acid (ATRA) has been reported to induce neuronal differentiation and cell-cycle arrest [Lopez-Carballo, Moreno, Masia, Perez, and Barettino (Activation of the phosphatidylinositol 3-kinase/Akt signalling pathway by retinoic acid is required for neural differentiation of SH-SY5Y human neuroblastoma cells. J Biol Chem 2002:277:25297-304.)]. In this study, we observed that ATRA-induced cellular ceramide accumulation, cell-growth arrest and differentiation accompanied with down-regulation of NCDase in SH-SY5Y cells, without a decrease in sphingosine or sphingosine 1-phosphate. We examined whether the down-regulation of NCDase was involved in the increase in ceramide and cell differentiation. ATRA was found to down-regulate mRNA, protein and the enzyme activity of NCDase. Interestingly, GATA-2 was also decreased with ATRA treatment, and experiments using its expression vector and siRNA and chromatin immunoprecipitation assay demonstrated GATA-2 acted as transcription-factor of NCDase gene expression. By establishing stable transfectants with decreased NCDase expression and activity, we clarified the significance of NCDase down-regulation for ATRA-induced neuronal differentiation. Those sub-clones showed both increased cellular ceramide and reduced cell growth as well as neuronal differentiation phenotypes. These results demonstrate that down-regulation of NCDase through ATRA-induced GATA-2 decrease plays an important role in induction of ceramide accumulation and neuronal differentiation in SH-SY5Y cells.",
author = "Kouji Tanaka and Keiko Tamiya-Koizumi and Kazumi Hagiwara and Hiromi Ito and Akira Takagi and Tetsuhito Kojima and Motoshi Suzuki and Soichiro Iwaki and Satoshi Fujii and Mitsuhiro Nakamura and Yoshiko Banno and Reiji Kannagi and Tatsuya Tsurumi and Mamoru Kyogashima and Takashi Murate",
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Tanaka, K, Tamiya-Koizumi, K, Hagiwara, K, Ito, H, Takagi, A, Kojima, T, Suzuki, M, Iwaki, S, Fujii, S, Nakamura, M, Banno, Y, Kannagi, R, Tsurumi, T, Kyogashima, M & Murate, T 2012, 'Role of down-regulated neutral ceramidase during all-trans retinoic acid-induced neuronal differentiation in SH-SY5Y neuroblastoma cells', Journal of Biochemistry, vol. 151, no. 6, pp. 611-620. https://doi.org/10.1093/jb/mvs033

Role of down-regulated neutral ceramidase during all-trans retinoic acid-induced neuronal differentiation in SH-SY5Y neuroblastoma cells. / Tanaka, Kouji; Tamiya-Koizumi, Keiko; Hagiwara, Kazumi; Ito, Hiromi; Takagi, Akira; Kojima, Tetsuhito; Suzuki, Motoshi; Iwaki, Soichiro; Fujii, Satoshi; Nakamura, Mitsuhiro; Banno, Yoshiko; Kannagi, Reiji; Tsurumi, Tatsuya; Kyogashima, Mamoru; Murate, Takashi.

In: Journal of Biochemistry, Vol. 151, No. 6, 01.06.2012, p. 611-620.

Research output: Contribution to journalArticle

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T1 - Role of down-regulated neutral ceramidase during all-trans retinoic acid-induced neuronal differentiation in SH-SY5Y neuroblastoma cells

AU - Tanaka, Kouji

AU - Tamiya-Koizumi, Keiko

AU - Hagiwara, Kazumi

AU - Ito, Hiromi

AU - Takagi, Akira

AU - Kojima, Tetsuhito

AU - Suzuki, Motoshi

AU - Iwaki, Soichiro

AU - Fujii, Satoshi

AU - Nakamura, Mitsuhiro

AU - Banno, Yoshiko

AU - Kannagi, Reiji

AU - Tsurumi, Tatsuya

AU - Kyogashima, Mamoru

AU - Murate, Takashi

PY - 2012/6/1

Y1 - 2012/6/1

N2 - Neutral ceramidase (NCDase) is considered to be a critical enzyme for controlling the turnover of ceramide, an important bioactive lipid, which determines cell's fate. All-trans retinoic acid (ATRA) has been reported to induce neuronal differentiation and cell-cycle arrest [Lopez-Carballo, Moreno, Masia, Perez, and Barettino (Activation of the phosphatidylinositol 3-kinase/Akt signalling pathway by retinoic acid is required for neural differentiation of SH-SY5Y human neuroblastoma cells. J Biol Chem 2002:277:25297-304.)]. In this study, we observed that ATRA-induced cellular ceramide accumulation, cell-growth arrest and differentiation accompanied with down-regulation of NCDase in SH-SY5Y cells, without a decrease in sphingosine or sphingosine 1-phosphate. We examined whether the down-regulation of NCDase was involved in the increase in ceramide and cell differentiation. ATRA was found to down-regulate mRNA, protein and the enzyme activity of NCDase. Interestingly, GATA-2 was also decreased with ATRA treatment, and experiments using its expression vector and siRNA and chromatin immunoprecipitation assay demonstrated GATA-2 acted as transcription-factor of NCDase gene expression. By establishing stable transfectants with decreased NCDase expression and activity, we clarified the significance of NCDase down-regulation for ATRA-induced neuronal differentiation. Those sub-clones showed both increased cellular ceramide and reduced cell growth as well as neuronal differentiation phenotypes. These results demonstrate that down-regulation of NCDase through ATRA-induced GATA-2 decrease plays an important role in induction of ceramide accumulation and neuronal differentiation in SH-SY5Y cells.

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