Transcriptional regulation of neutral sphingomyelinase 2 gene expression of a human breast cancer cell line, MCF-7, induced by the anti-cancer drug, daunorubicin

Hiromi Ito, Masashi Murakami, Ayako Furuhata, Siqiang Gao, Kayo Yoshida, Sayaka Sobue, Kazumi Hagiwara, Akira Takagi, Tetsuhito Kojima, Motoshi Suzuki, Yoshiko Banno, Kouji Tanaka, Keiko Tamiya-Koizumi, Mamoru Kyogashima, Yoshinori Nozawa, Takashi Murate

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Mg 2+ -dependent neutral SMases (NSMases) have emerged as prime candidates for stress-induced ceramide production. Among isoforms identified, previous reports have suggested the importance of NSMase2. However, its activation mechanism has not been precisely reported. Here, we analyzed the mechanism of NSMase2 gene expression by the anti-cancer drug, daunorubicin (DA). DA increased cellular ceramides (C16, C18 and C24) and NSMase activity of a human breast cancer cell line, MCF-7. DA remarkably increased the NSMase2 message and protein, whereas little change in NSMase1 and NSMase3 mRNAs and only a mild increase in acid SMase mRNA were observed. Overexpression and a knock down of NSMase2 indicated that NSMase2 played a role in DA-induced cell death. NSMase2 promoter analysis revealed that three Sp1 motifs located between -148 and -42bp upstream of the first exon were important in basic as well as in DA-induced promoter activity. Consistently, luciferase vectors containing three consensus Sp1-motifs but not its mutated form showed DA-induced transcriptional activation. DA-treated MCF-7 showed increased Sp3 protein. In SL2 cells lacking Sp family proteins, both Sp1 and Sp3 overexpression increased NSMase promoter activity. Increased binding of Sp family proteins by DA to three Sp1 motifs was shown by electrophoresis mobility shift and ChIP assays.

Original languageEnglish
Pages (from-to)681-690
Number of pages10
JournalBiochimica et Biophysica Acta - Gene Regulatory Mechanisms
Volume1789
Issue number11-12
DOIs
Publication statusPublished - 01-11-2009
Externally publishedYes

Fingerprint

Sphingomyelin Phosphodiesterase
Daunorubicin
Gene expression
Cells
Breast Neoplasms
Gene Expression
Cell Line
Pharmaceutical Preparations
Neoplasms
Proteins
Chemical activation
Messenger RNA
Ceramides
Electrophoretic Mobility Shift Assay
Cell death
Electrophoresis
Luciferases
Human Activities
Transcriptional Activation
Exons

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics

Cite this

Ito, Hiromi ; Murakami, Masashi ; Furuhata, Ayako ; Gao, Siqiang ; Yoshida, Kayo ; Sobue, Sayaka ; Hagiwara, Kazumi ; Takagi, Akira ; Kojima, Tetsuhito ; Suzuki, Motoshi ; Banno, Yoshiko ; Tanaka, Kouji ; Tamiya-Koizumi, Keiko ; Kyogashima, Mamoru ; Nozawa, Yoshinori ; Murate, Takashi. / Transcriptional regulation of neutral sphingomyelinase 2 gene expression of a human breast cancer cell line, MCF-7, induced by the anti-cancer drug, daunorubicin. In: Biochimica et Biophysica Acta - Gene Regulatory Mechanisms. 2009 ; Vol. 1789, No. 11-12. pp. 681-690.
@article{136678977d544bccab3cfb4d699c12ca,
title = "Transcriptional regulation of neutral sphingomyelinase 2 gene expression of a human breast cancer cell line, MCF-7, induced by the anti-cancer drug, daunorubicin",
abstract = "Mg 2+ -dependent neutral SMases (NSMases) have emerged as prime candidates for stress-induced ceramide production. Among isoforms identified, previous reports have suggested the importance of NSMase2. However, its activation mechanism has not been precisely reported. Here, we analyzed the mechanism of NSMase2 gene expression by the anti-cancer drug, daunorubicin (DA). DA increased cellular ceramides (C16, C18 and C24) and NSMase activity of a human breast cancer cell line, MCF-7. DA remarkably increased the NSMase2 message and protein, whereas little change in NSMase1 and NSMase3 mRNAs and only a mild increase in acid SMase mRNA were observed. Overexpression and a knock down of NSMase2 indicated that NSMase2 played a role in DA-induced cell death. NSMase2 promoter analysis revealed that three Sp1 motifs located between -148 and -42bp upstream of the first exon were important in basic as well as in DA-induced promoter activity. Consistently, luciferase vectors containing three consensus Sp1-motifs but not its mutated form showed DA-induced transcriptional activation. DA-treated MCF-7 showed increased Sp3 protein. In SL2 cells lacking Sp family proteins, both Sp1 and Sp3 overexpression increased NSMase promoter activity. Increased binding of Sp family proteins by DA to three Sp1 motifs was shown by electrophoresis mobility shift and ChIP assays.",
author = "Hiromi Ito and Masashi Murakami and Ayako Furuhata and Siqiang Gao and Kayo Yoshida and Sayaka Sobue and Kazumi Hagiwara and Akira Takagi and Tetsuhito Kojima and Motoshi Suzuki and Yoshiko Banno and Kouji Tanaka and Keiko Tamiya-Koizumi and Mamoru Kyogashima and Yoshinori Nozawa and Takashi Murate",
year = "2009",
month = "11",
day = "1",
doi = "10.1016/j.bbagrm.2009.08.006",
language = "English",
volume = "1789",
pages = "681--690",
journal = "Biochimica et Biophysica Acta - Gene Regulatory Mechanisms",
issn = "1874-9399",
publisher = "Elsevier",
number = "11-12",

}

Ito, H, Murakami, M, Furuhata, A, Gao, S, Yoshida, K, Sobue, S, Hagiwara, K, Takagi, A, Kojima, T, Suzuki, M, Banno, Y, Tanaka, K, Tamiya-Koizumi, K, Kyogashima, M, Nozawa, Y & Murate, T 2009, 'Transcriptional regulation of neutral sphingomyelinase 2 gene expression of a human breast cancer cell line, MCF-7, induced by the anti-cancer drug, daunorubicin', Biochimica et Biophysica Acta - Gene Regulatory Mechanisms, vol. 1789, no. 11-12, pp. 681-690. https://doi.org/10.1016/j.bbagrm.2009.08.006

Transcriptional regulation of neutral sphingomyelinase 2 gene expression of a human breast cancer cell line, MCF-7, induced by the anti-cancer drug, daunorubicin. / Ito, Hiromi; Murakami, Masashi; Furuhata, Ayako; Gao, Siqiang; Yoshida, Kayo; Sobue, Sayaka; Hagiwara, Kazumi; Takagi, Akira; Kojima, Tetsuhito; Suzuki, Motoshi; Banno, Yoshiko; Tanaka, Kouji; Tamiya-Koizumi, Keiko; Kyogashima, Mamoru; Nozawa, Yoshinori; Murate, Takashi.

In: Biochimica et Biophysica Acta - Gene Regulatory Mechanisms, Vol. 1789, No. 11-12, 01.11.2009, p. 681-690.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Transcriptional regulation of neutral sphingomyelinase 2 gene expression of a human breast cancer cell line, MCF-7, induced by the anti-cancer drug, daunorubicin

AU - Ito, Hiromi

AU - Murakami, Masashi

AU - Furuhata, Ayako

AU - Gao, Siqiang

AU - Yoshida, Kayo

AU - Sobue, Sayaka

AU - Hagiwara, Kazumi

AU - Takagi, Akira

AU - Kojima, Tetsuhito

AU - Suzuki, Motoshi

AU - Banno, Yoshiko

AU - Tanaka, Kouji

AU - Tamiya-Koizumi, Keiko

AU - Kyogashima, Mamoru

AU - Nozawa, Yoshinori

AU - Murate, Takashi

PY - 2009/11/1

Y1 - 2009/11/1

N2 - Mg 2+ -dependent neutral SMases (NSMases) have emerged as prime candidates for stress-induced ceramide production. Among isoforms identified, previous reports have suggested the importance of NSMase2. However, its activation mechanism has not been precisely reported. Here, we analyzed the mechanism of NSMase2 gene expression by the anti-cancer drug, daunorubicin (DA). DA increased cellular ceramides (C16, C18 and C24) and NSMase activity of a human breast cancer cell line, MCF-7. DA remarkably increased the NSMase2 message and protein, whereas little change in NSMase1 and NSMase3 mRNAs and only a mild increase in acid SMase mRNA were observed. Overexpression and a knock down of NSMase2 indicated that NSMase2 played a role in DA-induced cell death. NSMase2 promoter analysis revealed that three Sp1 motifs located between -148 and -42bp upstream of the first exon were important in basic as well as in DA-induced promoter activity. Consistently, luciferase vectors containing three consensus Sp1-motifs but not its mutated form showed DA-induced transcriptional activation. DA-treated MCF-7 showed increased Sp3 protein. In SL2 cells lacking Sp family proteins, both Sp1 and Sp3 overexpression increased NSMase promoter activity. Increased binding of Sp family proteins by DA to three Sp1 motifs was shown by electrophoresis mobility shift and ChIP assays.

AB - Mg 2+ -dependent neutral SMases (NSMases) have emerged as prime candidates for stress-induced ceramide production. Among isoforms identified, previous reports have suggested the importance of NSMase2. However, its activation mechanism has not been precisely reported. Here, we analyzed the mechanism of NSMase2 gene expression by the anti-cancer drug, daunorubicin (DA). DA increased cellular ceramides (C16, C18 and C24) and NSMase activity of a human breast cancer cell line, MCF-7. DA remarkably increased the NSMase2 message and protein, whereas little change in NSMase1 and NSMase3 mRNAs and only a mild increase in acid SMase mRNA were observed. Overexpression and a knock down of NSMase2 indicated that NSMase2 played a role in DA-induced cell death. NSMase2 promoter analysis revealed that three Sp1 motifs located between -148 and -42bp upstream of the first exon were important in basic as well as in DA-induced promoter activity. Consistently, luciferase vectors containing three consensus Sp1-motifs but not its mutated form showed DA-induced transcriptional activation. DA-treated MCF-7 showed increased Sp3 protein. In SL2 cells lacking Sp family proteins, both Sp1 and Sp3 overexpression increased NSMase promoter activity. Increased binding of Sp family proteins by DA to three Sp1 motifs was shown by electrophoresis mobility shift and ChIP assays.

UR - http://www.scopus.com/inward/record.url?scp=77949321187&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77949321187&partnerID=8YFLogxK

U2 - 10.1016/j.bbagrm.2009.08.006

DO - 10.1016/j.bbagrm.2009.08.006

M3 - Article

VL - 1789

SP - 681

EP - 690

JO - Biochimica et Biophysica Acta - Gene Regulatory Mechanisms

JF - Biochimica et Biophysica Acta - Gene Regulatory Mechanisms

SN - 1874-9399

IS - 11-12

ER -