TY - JOUR
T1 - New signalling pathway involved in the anti-proliferative action of vitamin D₃ and its analogues in human neuroblastoma cells. A role for ceramide kinase
AU - Bini, Francesca
AU - Frati, Alessia
AU - Garcia-Gil, Mercedes
AU - Battistini, Chiara
AU - Granado, Maria
AU - Martinesi, Maria
AU - Mainardi, Marco
AU - Vannini, Eleonora
AU - Luzzati, Federico
AU - Caleo, Matteo
AU - Peretto, Paolo
AU - Gomez-Muñoz, Antonio
AU - Meacci, Elisabetta
PY - 2012
Y1 - 2012
N2 - 1α,25-Dihydroxyvitamin D3 (1,25(OH)₂D₃), a crucial regulator of calcium/phosphorus homeostasis, has important physiological effects on growth and differentiation in a variety of malignant and non-malignant cells. Synthetic structural hormone analogues, with lower hypercalcemic side effects, are currently under clinical investigation. Sphingolipids appear to be crucial bioactive factors in the control of the cell fate: the phosphorylated forms, sphingosine-1-phosphate (S1P) and ceramide-1-phosphate (C1P), are mitogenic factors, whereas sphingosine and ceramide (Cer) usually act as pro-apoptotic agents. Although many studies correlate S1P function to impaired cell growth, the relevance of C1P/Cer system and its involvement in neuroblastoma cells remain to be clarified. Here, we demonstrated the anti-proliferative effect of 1,25(OH)₂D₃ as well as of its structural analogues, ZK156979 and ZK191784, in human SH-SY5Y cells, as judged by [³H]thymidine incorporation, cell growth and evaluation of active ERK1/2 levels. The inhibition of ceramide kinase (CerK), the enzyme responsible for C1P synthesis, by specific gene silencing or pharmacological inhibition, drastically reduced cell proliferation. 1,25(OH)₂D₃ and ZK191784 treatment induced a significant decrease in CerK expression and C1P content, and an increase of Cer. Notably, the treatment of SH-SY5Y cells with ZK159222, antagonist of 1,25(OH)₂D₃ receptor, trichostatin A, inhibitor of histone deacetylases, and COUP-TFI-siRNA prevented the decrease of CerK expression elicited by 1,25(OH)₂D₃ supporting the involvement of VDR/COUP-TFI/histone deacetylase complex in CerK regulation. Altogether, these findings provide the first evidence that CerK/C1P axis acts as molecular effector of the anti-proliferative action of 1,25(OH)₂D₃ and its analogues, thereby representing a new possible target for anti-cancer therapy of human neuroblastoma.
AB - 1α,25-Dihydroxyvitamin D3 (1,25(OH)₂D₃), a crucial regulator of calcium/phosphorus homeostasis, has important physiological effects on growth and differentiation in a variety of malignant and non-malignant cells. Synthetic structural hormone analogues, with lower hypercalcemic side effects, are currently under clinical investigation. Sphingolipids appear to be crucial bioactive factors in the control of the cell fate: the phosphorylated forms, sphingosine-1-phosphate (S1P) and ceramide-1-phosphate (C1P), are mitogenic factors, whereas sphingosine and ceramide (Cer) usually act as pro-apoptotic agents. Although many studies correlate S1P function to impaired cell growth, the relevance of C1P/Cer system and its involvement in neuroblastoma cells remain to be clarified. Here, we demonstrated the anti-proliferative effect of 1,25(OH)₂D₃ as well as of its structural analogues, ZK156979 and ZK191784, in human SH-SY5Y cells, as judged by [³H]thymidine incorporation, cell growth and evaluation of active ERK1/2 levels. The inhibition of ceramide kinase (CerK), the enzyme responsible for C1P synthesis, by specific gene silencing or pharmacological inhibition, drastically reduced cell proliferation. 1,25(OH)₂D₃ and ZK191784 treatment induced a significant decrease in CerK expression and C1P content, and an increase of Cer. Notably, the treatment of SH-SY5Y cells with ZK159222, antagonist of 1,25(OH)₂D₃ receptor, trichostatin A, inhibitor of histone deacetylases, and COUP-TFI-siRNA prevented the decrease of CerK expression elicited by 1,25(OH)₂D₃ supporting the involvement of VDR/COUP-TFI/histone deacetylase complex in CerK regulation. Altogether, these findings provide the first evidence that CerK/C1P axis acts as molecular effector of the anti-proliferative action of 1,25(OH)₂D₃ and its analogues, thereby representing a new possible target for anti-cancer therapy of human neuroblastoma.
KW - Antineoplastic Agents
KW - Calcitriol
KW - Cell Line, Tumor
KW - Cell Proliferation
KW - Cell Survival
KW - Ceramides
KW - Drugs, Investigational
KW - Enzyme Inhibitors
KW - Gene Silencing
KW - Histone Deacetylase Inhibitors
KW - Humans
KW - Molecular Targeted Therapy
KW - Neoplasm Proteins
KW - Neuroblastoma
KW - Phosphotransferases (Alcohol Group Acceptor)
KW - RNA, Small Interfering
KW - Receptors, Calcitriol
KW - Signal Transduction
KW - Vitamin D
KW - Antineoplastic Agents
KW - Calcitriol
KW - Cell Line, Tumor
KW - Cell Proliferation
KW - Cell Survival
KW - Ceramides
KW - Drugs, Investigational
KW - Enzyme Inhibitors
KW - Gene Silencing
KW - Histone Deacetylase Inhibitors
KW - Humans
KW - Molecular Targeted Therapy
KW - Neoplasm Proteins
KW - Neuroblastoma
KW - Phosphotransferases (Alcohol Group Acceptor)
KW - RNA, Small Interfering
KW - Receptors, Calcitriol
KW - Signal Transduction
KW - Vitamin D
UR - http://hdl.handle.net/10807/57487
U2 - 10.1016/j.neuropharm.2012.04.026
DO - 10.1016/j.neuropharm.2012.04.026
M3 - Article
SN - 0028-3908
VL - 63
SP - 524
EP - 537
JO - Neuropharmacology
JF - Neuropharmacology
ER -