title: Ceramide kinase contributes to proliferation but not to prostaglandin E2 formation in renal mesangial cells and fibroblasts.
abstract: OBJECTIVE
  Ceramide kinase (CerK) catalyzes the generation of the sphingolipid ceramide-1-phosphate (C1P) which regulates various cellular functions including cell growth and death, and inflammation. Here, we used a novel catalytic inhibitor of CerK, NVP-231, and CerK knockout cells to investigate the contribution of CerK to proliferation and inflammation in renal mesangial cells and fibroblasts.
  
  
  METHODS
  Cells were treated with NVP-231 and [(3)H]-thymidine incorporation into DNA, [(3)H]-arachidonic acid release, prostaglandin E2 (PGE2) synthesis, cell cycle distribution, and apoptosis were determined.
  
  
  RESULTS
  Treatment of rat mesangial cells and mouse renal fibroblasts with NVP-231 decreased DNA synthesis, but not of agonist-stimulated arachidonic acid release or PGE2 synthesis. Similarly, proliferation but not arachidonic acid release or PGE2 synthesis was reduced in CERK knockout renal fibroblasts. The anti-proliferative effect of NVP-231 on mesangial cells was due to M phase arrest as determined using the mitosis markers phospho-histone H3, cdc2 and polo-like kinase-1, and induction of apoptosis. Moreover, loss of CerK sensitized cells towards stress-induced apoptosis.
  
  
  CONCLUSIONS
  Our data demonstrate that CerK induces proliferation but not PGE2 formation of renal mesangial cells and fibroblasts, and suggest that targeted CerK inhibition has potential for treating mesangioproliferative kidney diseases.
authors: Institute of Pharmacology, University of Bern, Bern, Switzerland.
authors: Oleksandr
authors: O
authors: Pastukhov
authors: Stephanie
authors: S
authors: Schwalm
authors: Isolde
authors: I
authors: Römer
authors: Uwe
authors: U
authors: Zangemeister-Wittke
authors: Josef
authors: J
authors: Pfeilschifter
authors: Andrea
authors: A
authors: Huwiler
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D016718:Arachidonic Acid / Q000378:metabolism; D052160:Benzothiazoles / Q000494:pharmacology; D001952:Bridged-Ring Compounds / Q000494:pharmacology; D016203:CDC2 Protein Kinase / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D015232:Dinoprostone / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D055786:Gene Knockout Techniques; D006657:Histones / Q000378:metabolism; D007668:Kidney / Q000166:cytology*; D059566:M Phase Cell Cycle Checkpoints / Q000187:drug effects; D050527:Mesangial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D051381:Rats; D000097869:Polo-Like Kinase 1
chemical_list: D016718:Arachidonic Acid; D052160:Benzothiazoles; D001952:Bridged-Ring Compounds; D016203:CDC2 Protein Kinase; D018797:Cell Cycle Proteins; D015232:Dinoprostone; D006657:Histones; D017853:Phosphotransferases (Alcohol Group Acceptor); D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D000097869:Polo-Like Kinase 1; C534511:NVP 231; C066324:ceramide kinase