This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Okoshi, H. Articles by Igarashi, Y. Search for Related Content PubMed PubMed Citation Articles by Okoshi, H. Articles by Igarashi, Y.

Cell Membrane Signaling as Target in Cancer Therapy II: Inhibitory Effect of N,N,N-Trimethylsphingosine on Metastatic Potential of Murine B16 Melanoma Cell Line through Blocking of Tumor Cell-dependent Platelet Aggregation¹

The Biomembrane Institute, Seattle, Washington 98119; and Department of Pathobiology, University of Washington, Seattle, Washington 98195

Two phenotypic parameters, aberrant expression of protein kinase C and tumor cell-induced platelet aggregation (PA), have been correlated with abnormal growth behavior and metastatic potential of tumor cells. We recently observed that N,N,N-trimethylsphingosine (TMS) and N,N-dimethylsphingosine (DMS), but not sphingosine (SPN), had an inhibitory effect (via blocking of transmembrane signaling) on the growth of various human tumor cell lines in vitro as well as in vivo in nu/nu mice (K. Endo et al., Cancer Res., 51: 1613–1618, 1991). We therefore investigated the effects of TMS, DMS, and SPN on (a) PA induced by ADP and thrombin; (b) PA induced by melanoma cell line B16/BL6; and (c) experimental lung colonization as well as spontaneous lung metastasis of BL6 cells in syngeneic C57BL/6 mice. In experiments on agonist-induced PA, TMS inhibited PA and ATP secretion 5-fold more strongly than DMS or SPN. This effect may be based on the inhibition of M_r 47,000 platelet protein phosphorylation and/or inhibition of phosphatidylinositol turnover as a transmembrane signaling pathway in platelets. Tumor cell (BL6 melanoma)-induced PA and ATP secretion were also strongly inhibited by TMS, but not by DMS or SPN. Unlike ADP- or thrombin-induced PA, BL6 cell-induced PA was not inhibited by Calphostin-C (a potent protein kinase C inhibitor) or cilostazol (a potent inhibitor of PA based on inhibition of cyclic AMP phosphodiesterase). Since many previous studies suggested that the ability of tumor cells to induce PA is related to the degree of malignancy (e.g., metastatic potential) of tumor cells, we studied the effect of TMS on lung metastatic potential. Three independent sets of experiments, as described below, all showed clear inhibition of lung metastasis by administration of TMS: (a) i.v. coinjection of BL6 melanoma cells and TMS; (b) i.v. injection of TMS and, 1 h later, BL6 cells; (c) spontaneous metastasis to lung from s.c. BL6 tumor (TMS administered after establishment of tumor, followed by resection of tumor). In comparison to tumor growth inhibition produced by TMS or DMS, inhibition of melanoma metastasis by TMS is obvious at lower doses.

¹ This study was supported by funds from The Biomembrane Institute, in part under a research contract from Otsuka Pharmaceutical Co. (Osaka, Japan).

² Awardee, NCI Outstanding Investigator Grant CA42505.

³ To whom requests for reprints should be addressed, at The Biomembrane Institute, 201 Elliott Avenue West, Seattle, WA 98119.

Received 6/20/91. Accepted 9/ 5/91.

This article has been cited by other articles:

				K. Takabe, S. W. Paugh, S. Milstien, and S. Spiegel "Inside-Out" Signaling of Sphingosine-1-Phosphate: Therapeutic Targets Pharmacol. Rev., June 1, 2008; 60(2): 181 - 195. [Abstract] [Full Text] [PDF]

				S. Gundewar, J. W. Calvert, J. W. Elrod, and D. J. Lefer Cytoprotective effects of N,N,N-trimethylsphingosine during ischemia- reperfusion injury are lost in the setting of obesity and diabetes Am J Physiol Heart Circ Physiol, October 1, 2007; 293(4): H2462 - H2471. [Abstract] [Full Text] [PDF]

				T. Yoshimoto, M. Furuhata, S. Kamiya, M. Hisada, H. Miyaji, Y. Magami, K. Yamamoto, H. Fujiwara, and J. Mizuguchi Positive Modulation of IL-12 Signaling by Sphingosine Kinase 2 Associating with the IL-12 Receptor {beta}1 Cytoplasmic Region J. Immunol., August 1, 2003; 171(3): 1352 - 1359. [Abstract] [Full Text] [PDF]

				B. Campbell, Y. K. Shin, R. Scalia, and A. M. Lefer Beneficial effects of N,N,N-trimethylsphingosine following ischemia and reperfusion in the isolated perfused rat heart Cardiovasc Res, August 1, 1998; 39(2): 393 - 400. [Abstract] [Full Text] [PDF]

				T. Murohara, S. J. Parkinson, S. A. Waldman, and A. M. Lefer Inhibition of Nitric Oxide Biosynthesis Promotes P-selectin Expression in Platelets : Role of Protein Kinase C Arterioscler. Thromb. Vasc. Biol., November 1, 1995; 15(11): 2068 - 2075. [Abstract] [Full Text]

				S. M. Pitson, P. A. B. Moretti, J. R. Zebol, P. Xia, J. R. Gamble, M. A. Vadas, R. J. D'Andrea, and B. W. Wattenberg Expression of a Catalytically Inactive Sphingosine Kinase Mutant Blocks Agonist-induced Sphingosine Kinase Activation. A DOMINANT-NEGATIVE SPHINGOSINE KINASE J. Biol. Chem., October 20, 2000; 275(43): 33945 - 33950. [Abstract] [Full Text] [PDF]