Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
권호 표지
DOI QR코드

DOI QR Code

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
Download PDF
⟨ Previous Next ⟩

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.

Keywords

References

  1. Baeuerle, P. A. and D. Baltimore. 1996. NF-$\kappa$B: ten years after. 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 into the 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 antibodies to 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 human malignant melanoma: definition of six antigenic systems with 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 normal and 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. Enhancement of in vitro and in vivo anti-tumor activity of anti-GD2 monoclonal antibody 220-51 against human neuroblastoma by granulocyte-macrophage colony-stimulating factor and granulocyte colony-stimulating factor. Int. J. Mol. Med. 2, 471-475.
  8. Hakomori, S. 1996. Tumor malignancy defined by aberrant glycosylation 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 of GM3 $\alpha$-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 human CMP-NeuAc:GM3 $\alpha$2,8 sialyltransferase (GD3 synthase) gene through the NF-$\kappa$B activation in human melanoma SK-MEL-2 cells. Biochim. Biophys. Acta 1769, 622-630. https://doi.org/10.1016/j.bbaexp.2007.08.001
  12. Kang, N. Y., S. K Kang, Y. C. Lee, H. J. Choi, Y. S. Lee, S. Y. Cho, Y. S. Kim, J. H. Ko, and C. H. Kim. 2006. Transcriptional regulation of the human GD3 synthase gene expression in Fas-induced Jurkat T cells: a critical role of transcription factor NF-$\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-cell lymphoblastic 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, and Z. Peng. 2009. Apoptosis induced by synthetic retinoic acid CD437 on human melanoma A375 cells involves RIG-I pathway. 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. Lipid composition of human malignant melanoma tumors at various levels 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 ganglioside of human melanoma. Detection and characterisation by 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. Relationship of glycosyltransferases and mRNA levels to ganglioside expression in 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 of a 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 in melanoma 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, and A. N. Houghton. 1987. Stimulation of T lymphocyte proliferation by 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 human monoclonal 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-regulation in 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, and P. O. Livingston. 1998. Antibodies against GD2 ganglioside can eradicate syngeneic cancer micrometastases. Cancer Res. 58, 2844-2849.