High expression of sphingosine kinase 1 and S1P receptors in chemotherapy-resistant prostate cancer PC3 cells and their camptothecin-induced up-regulation

doi:10.1016/j.bbrc.2006.02.070

Biochemical and Biophysical Research Communications

Volume 342, Issue 4, 21 April 2006, Pages 1284-1290

https://doi.org/10.1016/j.bbrc.2006.02.070 Get rights and content

Abstract

Although most of pharmacological therapies for cancer utilize the apoptotic machinery of the cells, the available anti-cancer drugs are limited due to the ability of prostate cancer cells to escape from the anti-cancer drug-induced apoptosis. A human prostate cancer cell line PC3 is resistant to camptothecin (CPT). To elucidate the mechanism of this resistance, we have examined the involvement of sphingosine kinase (SPHK) and sphingosine 1-phosphate (S1P) receptor in CPT-resistant PC3 and -sensitive LNCaP cells. PC3 cells exhibited higher activity accompanied with higher expression levels of protein and mRNA of SPHK1, and also elevated expression of S1P receptors, S1P₁ and S1P₃, as compared with those of LNCaP cells. The knockdown of SPHK1 by small interfering RNA and inhibition of S1P receptor signaling by pertussis toxin in PC3 cells induced significant inhibition of cell growth, suggesting implication of SPHK1 and S1P receptors in cell proliferation in PC3 cells. Furthermore, the treatment of PC3 cells with CPT was found to induce up-regulation of the SPHK1/S1P signaling by induction of both SPHK1 enzyme and S1P₁/S1P₃ receptors. These findings strongly suggest that high expression and up-regulation of SPHK1 and S1P receptors protect PC3 cells from the apoptosis induced by CPT.

Section snippets

Materials and methods

Materials. [γ-³²P]ATP (3000 Ci/mmol) was from NEN Life Science Products (Boston, MA, USA). d-Erythro-sphingosine, N,N′-dimethylsphingosine (DMS) and sphingosine 1-phosphate (S1P) were purchased from Matreya (Pleassant Gap, PA), and pertussis toxin (PTX) was from Seikagaku (Tokyo, Japan). Camptothecin (CPT) was from Sigma (St. Louis, MO). Rabbit antibody against human SPHK1 (C-terminal peptide: CVEPPPSWKPQQMPPPEEPL) was generated in New Zealand White rabbits by injection with KLH-conjugated

SPHK1 activity, expression levels of protein and mRNA of SPHK1, and mRNA expression of S1P receptors in human prostate cancer cell lines PC3 and LNCaP

A number of evidence suggests that SPHK/S1P signaling is important for various cell functions including cell survival and proliferation [9], [10], [11], [12], [13]. To examine the difference in SPHK/S1P signaling between chemotherapy-resistant PC3 and -sensitive LNCaP cell lines, we examined the activity, protein and mRNA levels of SPHK1, and also the expression of S1P receptors, S1P_1–3 in these two cell lines. As shown in Fig. 1A, a higher level of sphingosine kinase (SPHK) activity was

Discussion

A number of studies have demonstrated that expression of SPHK1 would be linked to protection of cells from apoptosis [9], [10], [11], [12], [13]. Apoptosis is thought to be the predominant form of tumor cell demise caused by chemotherapy. Since resistance for anti-cancer drugs occurs often in cancer therapy, it is of interest to examine whether SPHK1 is implicated in chemotherapeutic resistance. It has been reported that overexpression of SPHK1 prevents anti-cancer drug-induced apoptosis in

Acknowledgments

This work was supported in part by Grants-in-Aid for Scientific Research (B) (09480162) and (C) (17510175) from the Ministry of Education, Science, Sports and Culture of Japan.

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High expression of sphingosine kinase 1 and S1P receptors in chemotherapy-resistant prostate cancer PC3 cells and their camptothecin-induced up-regulation

Abstract

Section snippets

Materials and methods

SPHK1 activity, expression levels of protein and mRNA of SPHK1, and mRNA expression of S1P receptors in human prostate cancer cell lines PC3 and LNCaP

Discussion

Acknowledgments

Trends Biochem. Sci.

Curr. Biol.

Exp. Cell Res.

Biochim. Biophys. Acta

Biochem. Biophys. Res. Commun.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Biologically active sphingolipids in cancer pathogenesis and treatment

Nat. Rev. Cancer

Sphingosine-1-phosphate: an enigmatic signalling lipid

Nat. Mol. Cell. Biol.

Functions of ceramide in coordinating cellular responses to stress

Science

Sphingolipid metabolism and cell growth regulation

FASEB J.

The therapeutic potential of modulating the ceramide/sphingomyelin pathway

J. Clin. Invest.

Sphingosine-1-phosphate as a second messenger in cell proliferation induced by PDGF and FCS mitogens

Nature