DNA damage induces down-regulation of UDP-glucose ceramide glucosyltransferase, increases ceramide levels and triggers apoptosis in p53-deficient cancer cells

Teka Ann S Haynes, Valery Filippov, Maria Filippova, Jun Yang, Kangling Zhang, Penelope J. Duerksen-Hughes

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

DNA damaging agents typically induce an apoptotic cascade in which p53 plays a central role. However, absence of a p53-mediated response does not necessarily abrogate programmed cell death, due to the existence of p53-independent apoptotic pathways, such as those mediated by the pro-apoptotic molecule ceramide. We compared ceramide levels before and after DNA damage in human osteosarcoma (U2OS) and colon cancer (HCT116) cells that were either expressing or deficient in p53. When treated with mitomycin C, p53-deficient cells, but not p53-expressing cells, showed a marked increase in ceramide levels. Microarray analysis of genes involved in ceramide metabolism identified acid ceramidase (ASAH1, up-regulated), ceramide glucosyltransferase (UGCG, down-regulated), and galactosylceramidase (GALC, up-regulated) as the three genes most affected. Experiments employing pharmacological and siRNA agents revealed that inhibition of UGCG is sufficient to increase ceramide levels and induce cell death. When inhibition of UGCG and treatment with mitomycin C were combined, p53-deficient, but not p53-expressing cells, showed a significant increase in cell death, suggesting that the regulation of sphingolipid metabolism could be used to sensitize cells to chemotherapeutic drugs.

Original languageEnglish (US)
Pages (from-to)943-953
Number of pages11
JournalBiochimica et Biophysica Acta - Molecular and Cell Biology of Lipids
Volume1821
Issue number7
DOIs
StatePublished - Jul 2012
Externally publishedYes

Fingerprint

ceramide glucosyltransferase
Ceramides
DNA Damage
Down-Regulation
Apoptosis
Cell Death
Mitomycin
Neoplasms
Acid Ceramidase
Galactosylceramidase
HCT116 Cells
Sphingolipids
Osteosarcoma
Microarray Analysis
Colonic Neoplasms
Small Interfering RNA
Genes
Pharmacology
DNA

Keywords

  • Apoptosis
  • Ceramide
  • Ceramide glucosyltransferase
  • HPV16 E6
  • p53

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

Cite this

DNA damage induces down-regulation of UDP-glucose ceramide glucosyltransferase, increases ceramide levels and triggers apoptosis in p53-deficient cancer cells. / Haynes, Teka Ann S; Filippov, Valery; Filippova, Maria; Yang, Jun; Zhang, Kangling; Duerksen-Hughes, Penelope J.

In: Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids, Vol. 1821, No. 7, 07.2012, p. 943-953.

Research output: Contribution to journalArticle

@article{b1b54aeae01e4fdd99c51b51b24f72c5,
title = "DNA damage induces down-regulation of UDP-glucose ceramide glucosyltransferase, increases ceramide levels and triggers apoptosis in p53-deficient cancer cells",
abstract = "DNA damaging agents typically induce an apoptotic cascade in which p53 plays a central role. However, absence of a p53-mediated response does not necessarily abrogate programmed cell death, due to the existence of p53-independent apoptotic pathways, such as those mediated by the pro-apoptotic molecule ceramide. We compared ceramide levels before and after DNA damage in human osteosarcoma (U2OS) and colon cancer (HCT116) cells that were either expressing or deficient in p53. When treated with mitomycin C, p53-deficient cells, but not p53-expressing cells, showed a marked increase in ceramide levels. Microarray analysis of genes involved in ceramide metabolism identified acid ceramidase (ASAH1, up-regulated), ceramide glucosyltransferase (UGCG, down-regulated), and galactosylceramidase (GALC, up-regulated) as the three genes most affected. Experiments employing pharmacological and siRNA agents revealed that inhibition of UGCG is sufficient to increase ceramide levels and induce cell death. When inhibition of UGCG and treatment with mitomycin C were combined, p53-deficient, but not p53-expressing cells, showed a significant increase in cell death, suggesting that the regulation of sphingolipid metabolism could be used to sensitize cells to chemotherapeutic drugs.",
keywords = "Apoptosis, Ceramide, Ceramide glucosyltransferase, HPV16 E6, p53",
author = "Haynes, {Teka Ann S} and Valery Filippov and Maria Filippova and Jun Yang and Kangling Zhang and Duerksen-Hughes, {Penelope J.}",
year = "2012",
month = "7",
doi = "10.1016/j.bbalip.2012.02.002",
language = "English (US)",
volume = "1821",
pages = "943--953",
journal = "Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids",
issn = "1388-1981",
publisher = "Elsevier",
number = "7",

}

TY - JOUR

T1 - DNA damage induces down-regulation of UDP-glucose ceramide glucosyltransferase, increases ceramide levels and triggers apoptosis in p53-deficient cancer cells

AU - Haynes, Teka Ann S

AU - Filippov, Valery

AU - Filippova, Maria

AU - Yang, Jun

AU - Zhang, Kangling

AU - Duerksen-Hughes, Penelope J.

PY - 2012/7

Y1 - 2012/7

N2 - DNA damaging agents typically induce an apoptotic cascade in which p53 plays a central role. However, absence of a p53-mediated response does not necessarily abrogate programmed cell death, due to the existence of p53-independent apoptotic pathways, such as those mediated by the pro-apoptotic molecule ceramide. We compared ceramide levels before and after DNA damage in human osteosarcoma (U2OS) and colon cancer (HCT116) cells that were either expressing or deficient in p53. When treated with mitomycin C, p53-deficient cells, but not p53-expressing cells, showed a marked increase in ceramide levels. Microarray analysis of genes involved in ceramide metabolism identified acid ceramidase (ASAH1, up-regulated), ceramide glucosyltransferase (UGCG, down-regulated), and galactosylceramidase (GALC, up-regulated) as the three genes most affected. Experiments employing pharmacological and siRNA agents revealed that inhibition of UGCG is sufficient to increase ceramide levels and induce cell death. When inhibition of UGCG and treatment with mitomycin C were combined, p53-deficient, but not p53-expressing cells, showed a significant increase in cell death, suggesting that the regulation of sphingolipid metabolism could be used to sensitize cells to chemotherapeutic drugs.

AB - DNA damaging agents typically induce an apoptotic cascade in which p53 plays a central role. However, absence of a p53-mediated response does not necessarily abrogate programmed cell death, due to the existence of p53-independent apoptotic pathways, such as those mediated by the pro-apoptotic molecule ceramide. We compared ceramide levels before and after DNA damage in human osteosarcoma (U2OS) and colon cancer (HCT116) cells that were either expressing or deficient in p53. When treated with mitomycin C, p53-deficient cells, but not p53-expressing cells, showed a marked increase in ceramide levels. Microarray analysis of genes involved in ceramide metabolism identified acid ceramidase (ASAH1, up-regulated), ceramide glucosyltransferase (UGCG, down-regulated), and galactosylceramidase (GALC, up-regulated) as the three genes most affected. Experiments employing pharmacological and siRNA agents revealed that inhibition of UGCG is sufficient to increase ceramide levels and induce cell death. When inhibition of UGCG and treatment with mitomycin C were combined, p53-deficient, but not p53-expressing cells, showed a significant increase in cell death, suggesting that the regulation of sphingolipid metabolism could be used to sensitize cells to chemotherapeutic drugs.

KW - Apoptosis

KW - Ceramide

KW - Ceramide glucosyltransferase

KW - HPV16 E6

KW - p53

UR - http://www.scopus.com/inward/record.url?scp=84860851547&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84860851547&partnerID=8YFLogxK

U2 - 10.1016/j.bbalip.2012.02.002

DO - 10.1016/j.bbalip.2012.02.002

M3 - Article

VL - 1821

SP - 943

EP - 953

JO - Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids

JF - Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids

SN - 1388-1981

IS - 7

ER -