Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Suppression of Human GD3 Synthase (hST8Sia I) Expression Induced by Retinoic Acid in Human Melanoma SK-MEL-2 Cells

흑색종세포주 SK-MEL-2에서 레티노이드에 의한 GD3합성효소(hST8Sia I)의 발현억제

  • Kwon, Haw-Young (Department of Biotechnology, College of Natural Resources and Life Science, Dong-A University) ;
  • Kang, Nam-Young (Department of Biotechnology, College of Natural Resources and Life Science, Dong-A University) ;
  • Lee, Young-Choon (Department of Biotechnology, College of Natural Resources and Life Science, Dong-A University)
  • 권화영 (동아대학교 생명자원과학대학 생명공학과) ;
  • 강남영 (동아대학교 생명자원과학대학 생명공학과) ;
  • 이영춘 (동아대학교 생명자원과학대학 생명공학과)
  • Received : 2010.02.08
  • Accepted : 2010.03.13
  • Published : 2010.05.31
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Abstract

To elucidate the mechanism underlying the suppressive regulation of hST8Sia I expression in retinoic acid (RA)-induced SK-MEL-2 cells, we characterized the promoter region of the hST8Sia I gene. Functional analysis of the 5‘-flanking region of the hST8Sia I gene by the transient expression method showed that the -1146 to -646 region, which contains putative binding sites for transcription factors c-Ets-1, CREB, AP-1 and NF-kB, functions as the RA-repressive promoter in SK-MEL-2 cells. Site-directed mutagenesis and ChIP analyses indicated that the NF-kB binding site at -731 to -722 is crucial for the RA-induced repression of hST8Sia I in SK-MEL-2 cells. In addition, the transcriptional activity of hST8Sia I suppressed by RA in SK-MEL-2 cells was strongly inhibited by extracellular signal-regulated protein kinase (ERK) inhibitor U0126 and protein kinase C (PKC) inhibitor GO6976, as determined by RT-PCR and luciferase assay of hST8Sia I promoter containing the -1146 to -646 regions. These results suggest that RA markedly modulates transcriptional regulation of hST8Sia I gene expression through the PKC/ERK signal pathway in SK-MEL-2 cells.

흑색종세포주 SK-MEL-2에서 레티노이드에 의한 GD3합성효소(hST8Sia I)의 발현억제기작을 규명하게 위하여 hST8Sia I의 프로모터 활성을 조사해 본 결과 -1146에서 -646영역에서 레티노이드에 의한 활성억제를 나타내었다. 또한 부위특이적 변이와 ChIP분석은 -731에서 -722영역에 위치한 전사인자NF-kB 결합부위가 hST8Sia I의 레티노이드에 의한 활성억제에 중요하게 관여하고 있음을 나타내었다. 이러한 발현 억제는 PKC/ERK 신호전달경로를 통하여 일어난다는 것을 신호전달경로 저해제를 이용한 RT-PCR과 프로모터 활성조사에 의해 규명하였다.

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References

  1. Baeuerle, P. A. and D. Baltimore. 1996. NF-$\kappa$B: ten yearsafter. Cell 87, 13-20. https://doi.org/10.1016/S0092-8674(00)81318-5
  2. Chen, F., V. Castranova, and X. Shi. 2001. New insights intothe role of nuclear factor-$\kappa$B in cell growth regulation. Am. J. Pathol. 159, 387-397. https://doi.org/10.1016/S0002-9440(10)61708-7
  3. Cheung, N. K., U. M. Saarinen, J. E. Neely, B. Landmeier,D. Donovan, and P. F. Coccia. 1985. Monoclonal antibodiesto a glycolipid antigen on human neuroblastoma cells.Cancer Res. 45, 2642-2649.
  4. Dippold, W. G., K. O. Lloyd, L. T. Li, H. Ikeda, H. F.Oettgen, and L. J. Old. 1980. Cell surface antigens of humanmalignant melanoma: definition of six antigenic systemswith mouse monoclonal antibodies. Proc. Natl. Acad. Sci. USA, 77, 6114-6118. https://doi.org/10.1073/pnas.77.10.6114
  5. Fields, A. L., D. R. Soprano, and K. J. Soprano. 2007.Retinoids in biological control and cancer. J. Cell Biochem.102, 886-898. https://doi.org/10.1002/jcb.21530
  6. Fligiel, S. E., D. R. Inman, H. S. Talwar, G. J. Fisher, J. J.Voorhees, and J. Varani. 1992. Modulation of growth in normaland malignant melanocytic cells by all-trans retinoic acid. J. Cutan Pathol. 19, 27-33. https://doi.org/10.1111/j.1600-0560.1992.tb01555.x
  7. Fukuda, M., K. Horibe, and K. Furukawa. 1998. Enhancementof in vitro and in vivo anti-tumor activity of anti-GD2 monoclonalantibody 220-51 against human neuroblastoma bygranulocyte-macrophage colony-stimulating factor andgranulocyte colony-stimulating factor. Int. J. Mol. Med. 2,471-475.
  8. Hakomori, S. 1996. Tumor malignancy defined by aberrantglycosylation and sphingo (glyco) lipid metabolism. Cancer Res. 56, 5309-5318.
  9. Hakomori, S. 1981.lycosphingolipids in cellular interaction,differentiation, and oncogenesis. Annu. Rev. Biochem. 50,733-764. https://doi.org/10.1146/annurev.bi.50.070181.003505
  10. Haraguchi, M., S. Yamashiro, A. Yamamoto, K. Furukawa,K. Takamiya, K. O. Lloyd, H. Shiku, and K. Furukawa. 1994.Isolation of GD3 synthase gene by expression cloning ofGM3 $\alpha$-2,8-sialyltransferase cDNA using anti-GD2 monoclonal antibody. Proc. Natl. Acad. Sci. USA 91, 10455-10459. https://doi.org/10.1073/pnas.91.22.10455
  11. Kang, N. Y., C. H. Kim, K. S. Kim, J. H. Ko, J. H. Lee,Y. K. Jeong, and Y. C. Lee. 2007. Expression of the humanCMP-NeuAc:GM3 $\alpha$2,8 sialyltransferase (GD3 synthase)gene through the NF-$\kappa$B activation in human melanomaSK-MEL-2 cells. Biochim. Biophys. Acta 1769, 622-630. https://doi.org/10.1016/j.bbaexp.2007.08.001
  12. Kang, N. Y., S. K Kang, Y. C. Lee, H. J. Choi, Y. S. Lee,S. Y. Cho, Y. S. Kim, J. H. Ko, and C. H. Kim. 2006.Transcriptional regulation of the human GD3 synthase geneexpression in Fas-induced Jurkat T cells: a critical role oftranscription factor NF-$\kappa$B in regulated expression.Glycobiology 16, 375-389. https://doi.org/10.1093/glycob/cwj087
  13. Merritt, W. D., J. T. Casper, S. J. Lauer and G. H. Reaman.1987. Expression of GD3 ganglioside in childhood T-celllymphoblastic malignancies. Cancer Res. 47, 1724-1730.
  14. Old, L. J. 1981. Cancer immunology: the search for specificity-G. H. A. Clowes Memorial lecture. Cancer Res. 41,361-375.
  15. Pan, M., S. Geng, S. Xiao, J. Ren, Y. Liu, X. Li, Z. Li, andZ. Peng. 2009. Apoptosis induced by synthetic retinoic acidCD437 on human melanoma A375 cells involves RIG-Ipathway. Arch. Dermatol. Res. 301, 15-20. https://doi.org/10.1007/s00403-008-0902-x
  16. Portoukalian, J., G. Zwingelstein, and J. F. Dore. 1979. Lipidcomposition of human malignant melanoma tumors at variouslevels of malignant growth. Eur. J. Biochem. 94, 19-23. https://doi.org/10.1111/j.1432-1033.1979.tb12866.x
  17. Pukel, C. S., K. O. Lloyd, L. R. Travassos, W. G. Dippold,H. F. Oettgen, and L. J. Old. 1982. GD3, a prominent gangliosideof human melanoma. Detection and characterisationby mouse monoclonal antibody. J. Exp. Med. 155,1133-1147. https://doi.org/10.1084/jem.155.4.1133
  18. Rusan, S., B. K. Raj, and K. O. Lloyd. 1999. Relationshipof glycosyltransferases and mRNA levels to ganglioside expressionin neuroblastoma and melanoma cells. J. Neurochem.72, 514-521. https://doi.org/10.1046/j.1471-4159.1999.0720514.x
  19. Sasaki, K., K. Kurata, N. Kojima, N. Kurosawa, S. Ohta, N.Hanai, S. Tsuji, and T. Nishi. 1994. Expression cloning ofa GM3-specific $\alpha$-2,8-sialyltransferase (GD3 synthase). J. Biol. Chem. 269, 15950-15956.
  20. Sporn, M. B. and A. B. Roberts. 1983. Role of retinoids in differentiation and carcinogenesis, Cancer Res. 43, 3034-3040.
  21. Svennerholm, L. 1980. Gangliosides and synaptic transmission.Adv. Exp. Med. Biol. 125, 533-544. https://doi.org/10.1007/978-1-4684-7844-0_46
  22. Thampoe, I. J., K. Furukawa, E. Vellvé, and K. O. Lloyd.1989. Sialyltransferase levels and ganglioside expression inmelanoma and other cultured human cancer cells. Cancer Res. 49, 6258-6264.
  23. Welte, K., G. Miller, P. B. Chapman, H. Yuasa, E. Natoli,J. E. Kunicka, C. Cordon-Cardo, C. Buhrer, L. J. Old, andA. N. Houghton. 1987. Stimulation of T lymphocyte proliferationby monoclonal antibodies against GD3 ganglioside.J. Immunol. 139, 1763-1771.
  24. Yamaguchi, H., K. Furukawa, S. R. Fortunato, P. O. Livingston, K. O. Lloyd, H. F. Oettgen, and L. J. Old. 1987.Cell-surface antigens of melanoma recognized by humanmonoclonal antibodies. Proc. Natl. Acad. Sci. USA 84,2416-2420. https://doi.org/10.1073/pnas.84.8.2416
  25. Yamashiro, S., M. Okada, M. Haraguchi, K. Furukawa, K.O. Lloyd, H. Shiku, and K. Furukawa. 1995. Expression of$\alpha$2,8-sialyltransferase (GD3 synthase) gene in human cancer cell lines: high level expression in melanomas and up-regulationin activated T lymphocytes. Glycoconj. J. 12, 894-900. https://doi.org/10.1007/BF00731251
  26. Zhang, H., S. Zhang, N.-K. V. Cheung, G. Ragupathi, andP. O. Livingston. 1998. Antibodies against GD2 gangliosidecan eradicate syngeneic cancer micrometastases. Cancer Res.58, 2844-2849.