Cell density-dependent reduction of dihydroceramide desaturase activity in neuroblastoma cells

Stefka D. Spassieva, Mehrdad Rahmaniyan, Jacek Bielawski, Christopher J. Clarke, Jacqueline M. Kraveka, Lina M. Obeid

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

We applied a metabolic approach to investigate the role of sphingolipids in cell density-induced growth arrest in neuroblastoma cells. Our data revealed that sphingolipid metabolism in neuroblastoma cells significantly differs depending on the cells' population context. At high cell density, cells exhibited G0/G1 cell-cycle arrest and reduced ceramide, monohexosylceramide, and sphingomyelin, whereas dihydroceramide was significantly increased. In addition, our metabolic-labeling experiments showed that neuroblastoma cells at high cell density preferentially synthesized very long chain (VLC) sphingolipids and dramatically decreased synthesis of sphingosine-1-phosphate (S1P). Moreover, densely populated neuroblastoma cells showed increased message levels of both anabolic and catabolic enzymes of the sphingolipid pathway. Notably, our metabolic-labeling experiments indicated reduced dihydroceramide desaturase activity at confluence, which was confirmed by direct measurement of dihydroceramide desaturase activity in situ and in vitro. Importantly, we could reduce dihydroceramide desaturase activity in low-density cells by applying conditional media from high-density cells, as well as by adding reducing agents, such as DTT and L -cysteine to the media. In conclusion, our data suggest a role of the sphingolipid pathway, dihydroceramides desaturase in particular, in confluence-induced growth arrest in neuroblastoma cells.

Original languageEnglish
Pages (from-to)918-928
Number of pages11
JournalJournal of Lipid Research
Volume53
Issue number5
DOIs
StatePublished - May 2012

Keywords

  • Cell-cycle arrest
  • Ceramide
  • Dihydroceramide
  • Sphingolipid

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Cell Biology

Fingerprint

Dive into the research topics of 'Cell density-dependent reduction of dihydroceramide desaturase activity in neuroblastoma cells'. Together they form a unique fingerprint.

Cite this