Implications of sphingosine kinase 1 expression level for the cellular sphingolipid rheostat: relevance as a marker for daunorubicin sensitivity of leukemia cells
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We recently reported increased sphingosine kinase 1 (SPHK1) and decreased neutral sphingomyelinase 2 (NSMase2) gene expression in myelodysplastic syndromes and acute leukemia. This alteration is supposed to change the cellular sphingolipid metabolites; however, positive correlations were observed between daunorubicin (DA)-IC50 and the SPHK1 message but not between DA-IC50 and NSMase2 messages, when 16 different leukemia cell lines were used to analyze the relationship between gene expressions and chemosensitivity against DA. Using two cell lines with either the highest or lowest SPHK1 expression, cellular ceramides and sphingosine 1-phosphate (S1P) were quantified by liquid chromatography/mass spectrometry. Increased ceramide was observed in DA-sensitive, but not in DA-resistant cell lines treated with low doses of DA. Upon DA treatment, S1P decreased more in the sensitive cell lines than in resistant cell lines. A SPHK inhibitor recovered the DA sensitivity of DA-resistant cells. The modulation of SPHK1 gene expression by either overexpression or using siRNA affected the DA sensitivity of representative cell lines. Results clearly show that SPHK1 is both a good marker to predict the DA sensitivity of leukemia cells and a potential therapeutic target for leukemia with high SPHK1 expression, and suggest that the sphingolipid rheostat plays a significant role in DA-induced cytotoxicity.
KeywordsSphingosine kinase 1 Daunorubicin Chemosensitivity Ceramide Sphingosine 1-phosphate
- C16 ceramide
- C18 ceramide
- C24 ceramide
Reverse transcription-polymerase chain reaction
Liquid chromatography-tandem mass spectrometry
Electrospray atmospheric pressure ionization
Multiple reaction monitoring
The authors express their sincere thanks to Dr. H. Nagai, Ms. K. Hagiwara (Research Center for Blood Diseases, National Hospital Organization Nagoya Medical Center, Nagoya, Japan), Dr. S.M. Pitson (University of Adelaide, Australia), and Dr. Y.A. Hannun (University of South Carolina, SC, USA) for providing leukemia cell lines and expression vectors. We also express our gratitude to Dr. M. Kyogashima and Dr. K. Koizumi-T. (Aichi Cancer Center, Nagoya, Japan) for their assistance with the ceramide quantification.