title: The ceramide kinase inhibitor NVP-231 inhibits breast and lung cancer cell proliferation by inducing M phase arrest and subsequent cell death.
abstract: OBJECTIVE
  Ceramide kinase (CerK) catalyzes the generation of ceramide-1-phosphate which may regulate various cellular functions, including inflammatory reactions and cell growth. Here, we studied the effect of a recently developed CerK inhibitor, NVP-231, on cancer cell proliferation and viability and investigated the role of cell cycle regulators implicated in these responses.
  
  
  METHODS
  The breast and lung cancer cell lines MCF-7 and NCI-H358 were treated with increasing concentrations of NVP-231 and DNA synthesis, colony formation and cell death were determined. Flow cytometry was performed to analyse cell cycle distribution of cells and Western blot analysis was used to detect changes in cell cycle regulator expression and activation.
  
  
  RESULTS
  In both cell lines, NVP-231 concentration-dependently reduced cell viability, DNA synthesis and colony formation. Moreover it induced apoptosis, as measured by increased DNA fragmentation and caspase-3 and caspase-9 cleavage. Cell cycle analysis revealed that NVP-231 decreased the number of cells in S phase and induced M phase arrest with an increased mitotic index, as determined by increased histone H3 phosphorylation. The effect on the cell cycle was even more pronounced when NVP-231 treatment was combined with staurosporine. Finally, overexpression of CerK protected, whereas down-regulation of CerK with siRNA sensitized, cells for staurosporine-induced apoptosis.
  
  
  CONCLUSIONS
  Our data demonstrate for the first time a crucial role for CerK in the M phase control in cancer cells and suggest its targeted inhibition, using drugs such as NVP-231, in combination with conventional pro-apoptotic chemotherapy.
authors.affiliation: Institute of Pharmacology, University of Bern, Bern, Switzerland.
authors.forename: Oleksandr
authors.initials: O
authors.lastname: Pastukhov
authors.forename: Stephanie
authors.initials: S
authors.lastname: Schwalm
authors.forename: Uwe
authors.initials: U
authors.lastname: Zangemeister-Wittke
authors.forename: Doriano
authors.initials: D
authors.lastname: Fabbro
authors.forename: Frederic
authors.initials: F
authors.lastname: Bornancin
authors.forename: Lukasz
authors.initials: L
authors.lastname: Japtok
authors.forename: Burkhard
authors.initials: B
authors.lastname: Kleuser
authors.forename: Josef
authors.initials: J
authors.lastname: Pfeilschifter
authors.forename: Andrea
authors.initials: A
authors.lastname: Huwiler
mesh_terms: D000230:Adenocarcinoma; D017209:Apoptosis / Q000187:drug effects*; D052160:Benzothiazoles / Q000494:pharmacology*; D015153:Blotting, Western; D001943:Breast Neoplasms; D001952:Bridged-Ring Compounds / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival; D004247:DNA / Q000096:biosynthesis / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005434:Flow Cytometry; D006801:Humans; D008175:Lung Neoplasms; D059566:M Phase Cell Cycle Checkpoints / Q000187:drug effects*; D061986:MCF-7 Cells; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000037:antagonists & inhibitors*; D019311:Staurosporine / Q000494:pharmacology; D014410:Tumor Stem Cell Assay
chemical_list: D052160:Benzothiazoles; D001952:Bridged-Ring Compounds; D004791:Enzyme Inhibitors; C534511:NVP 231; D004247:DNA; D017853:Phosphotransferases (Alcohol Group Acceptor); C066324:ceramide kinase; D019311:Staurosporine