Ceramide kinase contributes to proliferation but not to prostaglandin E2 formation in renal mesangial cells and fibroblasts.

Abstract

BACKGROUND/AIMS:

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: Oleksandr Pastukhov, Stephanie Schwalm, Isolde Römer, Uwe Zangemeister-Wittke, Josef Pfeilschifter, Andrea Huwiler
• Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology (Cell Physiol Biochem) Vol. 34 Issue 1 Pg. 119-33 ( 2014) ISSN: 1421-9778 [Electronic] Germany
• PMID: 24977486 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Benzothiazoles
• Bridged-Ring Compounds
• Cell Cycle Proteins
• Histones
• NVP 231
• Proto-Oncogene Proteins
• Arachidonic Acid
• Phosphotransferases (Alcohol Group Acceptor)
• ceramide kinase
• Protein Serine-Threonine Kinases
• polo-like kinase 1
• CDC2 Protein Kinase
• Dinoprostone

Topics

• Animals
• Apoptosis (drug effects)
• Arachidonic Acid (metabolism)
• Benzothiazoles (pharmacology)
• Bridged-Ring Compounds (pharmacology)
• CDC2 Protein Kinase (metabolism)
• Cell Cycle Proteins (metabolism)
• Cell Proliferation (drug effects)
• Cells, Cultured
• Dinoprostone (metabolism)
• Fibroblasts (cytology, drug effects, metabolism)
• Gene Knockout Techniques
• Histones (metabolism)
• Kidney (cytology)
• M Phase Cell Cycle Checkpoints (drug effects)
• Mesangial Cells (cytology, drug effects, metabolism)
• Mice
• Mice, Inbred BALB C
• Phosphotransferases (Alcohol Group Acceptor) (deficiency, genetics, metabolism)
• Protein Serine-Threonine Kinases (metabolism)
• Proto-Oncogene Proteins (metabolism)
• Rats