We found that natural (long-chain) ceramide 1-phosphate can be dispersed into aqueous solution when dissolved in an appropriate mixture of methanol/dodecane (49:1, v/v). This solvent mixture facilitates the interaction of this phosphosphingolipid with cells. Under these conditions, incubation of EGFR T17 fibroblasts with natural ceramide 1-phosphate caused a potent stimulation of DNA synthesis. This effect was accompanied by an increase in the levels of proliferating-cell nuclear antigen. Concentrations of natural ceramide 1-phosphate that stimulated the synthesis of DNA did not inhibit adenylate cyclase activity, nor did they stimulate phospholipase D. Natural ceramide 1-phosphate did not alter the cellular phosphorylation state of tyrosine residues or of mitogen-activated protein kinase. Furthermore, natural ceramide 1-phosphate failed to induce the expression of the proto-oncogenes c-myc and c-fos. Both the stimulation of DNA synthesis and the induction of proliferating-cell nuclear antigen by natural ceramide 1-phosphate were inhibited by natural ceramides. This work suggests that the use of methanol and dodecane to deliver natural ceramide 1-phosphate to cells may be useful for elucidation of the biological function(s) and mechanism(s) of action of ceramide 1-phosphate.
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July 1997
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Research Article|
July 15 1997
Stimulation of DNA synthesis by natural ceramide 1-phosphate
Antonio GOMEZ-MUÑOZ;
1Instituto de Investigaciones Biomédicas, Consejo Superior de Investigaciones Cientéficas (CSIC), C/Arturo Duperier 4, 28029-Madrid, Spain (Instituto Asociado al Grupo de Biomembranas de la Facultad de Ciencias de la Universidad del Paés Vasco, P.O. Box 644, Bilbao, Spain)
†To whom correspondence should be addressed.
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Laura M. FRAGO;
Laura M. FRAGO
*
1Instituto de Investigaciones Biomédicas, Consejo Superior de Investigaciones Cientéficas (CSIC), C/Arturo Duperier 4, 28029-Madrid, Spain (Instituto Asociado al Grupo de Biomembranas de la Facultad de Ciencias de la Universidad del Paés Vasco, P.O. Box 644, Bilbao, Spain)
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Luis ALVAREZ;
Luis ALVAREZ
1Instituto de Investigaciones Biomédicas, Consejo Superior de Investigaciones Cientéficas (CSIC), C/Arturo Duperier 4, 28029-Madrid, Spain (Instituto Asociado al Grupo de Biomembranas de la Facultad de Ciencias de la Universidad del Paés Vasco, P.O. Box 644, Bilbao, Spain)
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Isabel VARELA-NIETO
Isabel VARELA-NIETO
1Instituto de Investigaciones Biomédicas, Consejo Superior de Investigaciones Cientéficas (CSIC), C/Arturo Duperier 4, 28029-Madrid, Spain (Instituto Asociado al Grupo de Biomembranas de la Facultad de Ciencias de la Universidad del Paés Vasco, P.O. Box 644, Bilbao, Spain)
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Publisher: Portland Press Ltd
Received:
October 15 1996
Revision Received:
February 14 1997
Accepted:
March 11 1997
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1997
1997
Biochem J (1997) 325 (2): 435–440.
Article history
Received:
October 15 1996
Revision Received:
February 14 1997
Accepted:
March 11 1997
Citation
Antonio GOMEZ-MUÑOZ, Laura M. FRAGO, Luis ALVAREZ, Isabel VARELA-NIETO; Stimulation of DNA synthesis by natural ceramide 1-phosphate. Biochem J 15 July 1997; 325 (2): 435–440. doi: https://doi.org/10.1042/bj3250435
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