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Journal of Biochemistry Advance Access published online on October 25, 2008
Journal of Biochemistry, doi:10.1093/jb/mvn138
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© 2008 The Japanese Biochemical Society

Regulation by Sphingolipids of the Fate of FRTL-5 Cells

Yumiko Satoh*, Xin Li*, Hiromitsu Yokota, Makoto Osada, Yukio Ozaki, Ryohei Katoh and Yutaka Yatomi

Department of Clinical Laboratory, The University of Tokyo Hospital, Tokyo, Japan.
Department of Pathology, Faculty of Medicine, University of Yamanashi, Yamanashi, Japan
Department of Clinical and Laboratory Medicine, Faculty of Medicine, University of Yamanashi, Yamanashi, Japan
Department of Clinical Laboratory Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan

Correspondence to: Yutaka Yatomi, MD, PhD, Department of Clinical Laboratory Medicine, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan. Tel: +81-3-5800-8721; Fax: +81-3-5689-0495; E-mail: yatoyuta-tky{at}umin.ac.jp

Received August 4, 2008; Accepted October 6, 2008


  Abstract

Sphingolipids, including ceramide (Cer), sphingosine (Sph), and sphingosine 1-phosphate (Sph-1-P) have recently emerged as signal-transducing molecules. Functionally, a distinguishing characteristic of these lipids is their apparent participation in pro- or anti-proliferative cell regulation pathways. In this study, we examined the involvement of sphingolipids in the fate of FRTL-5 thyroid follicular cells. We first examined the effects of sphingolipids on FRTL-5 cell viability. Sph and Cer induced apoptosis, as revealed by fluorescence microscopy of TUNEL-positive fragmented nuclei and 180-300 bp DNA fragmentation on agarose gel electrophoresis while Sph-1-P was confirmed to prevent FRTL-5 cell apoptosis induced by deprivation of serum and TSH, possibly via cell surface receptors. We then analyzed the metabolism of radiolabeled Sph and C6-Cer (a synthetic cell-permeable Cer) in FRTL-5 cells by thin layer chromatography, followed by autoradiography. Sph was mainly metabolized to Cer, and then to sphingomyelin, while Sph conversion into Sph-1-P was hardly detected. These changes were not affected by stimulation of the cells with TSH. Our results indicate the involvement of sphingolipid mediators in the fate of FRTL-5 thyroid cells.

Key Words: ceramide, FRTL-5, sphingosine, sphingosine 1-phosphate, thyrocyte

*These two authors equally contributed to this work.


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