Increasing intracellular ceramide: an approach that enhances the cytotoxic response in prostate cancer cells.

摘要

OBJECTIVES: To investigate the feasibility of targeting ceramide metabolism to enhance chemotherapy cytotoxicity in prostate cancer. Discovering new targets for cancer treatment is an important endeavor, especially in prostate malignancies, which often revert to hormone- and chemotherapy-refractory disease states. METHODS: Ceramide metabolism was measured in human prostate cancer cell lines using [(3)H]palmitic acid as the tracer. Cellular lipids were analyzed by thin-layer chromatography and liquid scintillation counting. Cell viability in response to drug exposure was measured spectrophotometrically using commercial cell proliferation reagents. RESULTS: LNCaP cells were five times more sensitive to N-(4-hydroxyphenyl)retinamide (4-HPR), a synthetic retinoid, compared with PC-3 cells. Ceramide levels increased only twofold in PC-3 cells versus 10-fold in LNCaP cells in response to 10 microM 4-HPR. PC-3 resistance to 4-HPR could be reversed by the addition of tamoxifen or other agents that block the metabolism of ceramide to glucosylceramide, and with tamoxifen this was marked by a ninefold increase in cellular ceramide levels. The influence of 4-HPR on ceramide metabolism was shown to be through activation of serine palmitoyltransferase, the rate-limiting enzyme in the ceramide synthesis pathway. Blocking the ceramide generated by 4-HPR reduced the extent of apoptosis. CONCLUSIONS: Increasing intracellular concentrations of ceramide may be an avenue to enhance the cytotoxic response to chemotherapy in human prostate cancer.