| Downregulation of neutral ceramidase by gemcitabine: Implications for cell cycle regulation. | |
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MedLine Citation:
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PMID: 19345744 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Gemcitabine (GMZ) is a chemotherapeutic agent with well established effects on cell growth arrest and apoptosis. In this study, we investigated the potential roles of bioactive sphingolipids in mediating the growth suppressing effects of GMZ on a polyoma middle T transformed murine endothelial cell line. After 12-hour GMZ (0.6 microM) treatment, cell growth was arrested at the G(0)/G(1) phase as detected by flow cytometric cell cycle analysis and MTT cell viability analysis, and this was accompanied by dephosphorylation of the retinoblastoma protein (Rb). Furthermore, GMZ treatment resulted in increased levels of specifically the very long chain ceramides as determined by mass spectrometry. Mechanistically, GMZ did not appear to affect the activities of many enzymes of ceramide metabolism; however, GMZ caused a selective reduction in the protein levels of neutral ceramidase (NCDase), as indicated by Western blot analysis, with a concomitant decrease in NCDase activity. The significance of NCDase loss on cell cycle regulation was investigated by specific knockdown of the enzyme using small interfering RNA (siRNA). Interestingly, NCDase siRNA transfection was sufficient to induce a cell cycle arrest at G(0)/G(1) and an increase in total ceramide levels, with significant elevation in very long chain ceramides (C(24:1) and C(24:0)). NCDase siRNA also induced Rb dephosphorylation. These data provide evidence for a novel mechanism of action for GMZ and highlight downregulation of NCDase as a critical step in GMZ-mediated ceramide elevation and cell cycle arrest. |
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Authors:
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Bill X Wu; Youssef H Zeidan; Yusuf A Hannun |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural Date: 2009-04-02 |
Journal Detail:
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Title: Biochimica et biophysica acta Volume: 1791 ISSN: 0006-3002 ISO Abbreviation: Biochim. Biophys. Acta Publication Date: 2009 Aug |
Date Detail:
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Created Date: 2009-07-20 Completed Date: 2009-09-23 Revised Date: 2010-12-03 |
Medline Journal Info:
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Nlm Unique ID: 0217513 Medline TA: Biochim Biophys Acta Country: Netherlands |
Other Details:
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Languages: eng Pagination: 730-9 Citation Subset: IM |
Affiliation:
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Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, 29425, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cell Cycle / drug effects* Cell Line Ceramides / metabolism Deoxycytidine / analogs & derivatives*, pharmacology Down-Regulation / drug effects* Endothelial Cells / cytology*, drug effects, enzymology* Gene Knockdown Techniques Mice Neutral Ceramidase / metabolism* Phosphorylation / drug effects RNA, Small Interfering / metabolism Retinoblastoma Protein / metabolism Sphingomyelin Phosphodiesterase / metabolism |
| Grant Support | |
ID/Acronym/Agency:
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CA87584/CA/NCI NIH HHS; GM43825/GM/NIGMS NIH HHS; P01 CA097132-06/CA/NCI NIH HHS; R01 CA087584-09/CA/NCI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Ceramides; 0/RNA, Small Interfering; 0/Retinoblastoma Protein; 103882-84-4/gemcitabine; 951-77-9/Deoxycytidine; EC 3.1.4.12/Sphingomyelin Phosphodiesterase; EC 3.5.1.23/Neutral Ceramidase |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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