Suppressive activity of protease inhibitors from buckwheat seeds against human T-acute lymphoblastic leukemia cell lines

Original Articles

Abstract

The buckwheat protease inhibitor designated BWI-1, a member of the potato inhibitor I family, inhibits trypsin, chymotrypsin, and subtilisin, whereas the buckwheat protease inhibitor designated BWI-2a, a novel protease inhibitor homologous to the vicilin family, inhibits only trypsin. We examined the suppressive activity of BWI-1 and BWI-2a against T-acute lymphoblastic leukemia (T-ALL) cells, such as JURKAT and CCRF-CEM, and human normal blood lymphocytes. Both inhibitors significantly suppressed the growth of T-ALL cells as judged by the soluble 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (tetrazolium/formazan assay). JURKAT cells showed slightly higher susceptibility to buckwheat inhibitors than CCRF-CEM cells. Modification of Arg residue(s) in inhibitors by 1,2-cyclohexandione inactivated their trypsin inhibitory activity, considerably abolishing their suppressive activity. This suggests that the trypsin inhibitory activity is involved in the suppression of growth of human T-ALL cell lines. It was further found that both inhibitors triggered programmed cell death (apoptosis) of these cell strains with DNA fragmentation.

Index Entries

Apoptosis buckwheat protease inhibitor tumor hematopoietic stem cell T-acute lymphoblastic leukemia 

References

  1. 1.
    Bode, W. and Huber, R. (1992), Eur. J. Biochem. 204, 433–451.PubMedCrossRefGoogle Scholar
  2. 2.
    Troll, W. and Yavelow, J. (1983), in Basic Research to Policy Implications, Roe D. A., ed., Alan R. Liss, New York, pp. 167–176.Google Scholar
  3. 3.
    Yavelow, J., Finlay, T. H., Kennedy, A. R., and Troll, W. (1983), Cancer Res. 43, 2454s-2459s.PubMedGoogle Scholar
  4. 4.
    Yavelow, J., Collins, M., Birk, Y., Troll, W., and Kennedy, A. R. (1985), Proc. Natl. Acad. Sci. USA 82, 5395–5399.PubMedCrossRefADSGoogle Scholar
  5. 5.
    Lin, J.-Y., Hsieh, Y.-S., and Chu, S.-C. (1989), Biochem. Int. 19, 313–323.PubMedGoogle Scholar
  6. 6.
    Ohba, H., Nishikawa, M., Kimura, M., Yamasaki, N., and Moriwaki, S. (1998), Biosci. Biotechnol. Biochem. 62, 1166–1170.PubMedCrossRefGoogle Scholar
  7. 7.
    Park, S.-S., Abe, K., Kimura, M., Urisu, A., and Yamasaki, N. (1997), FEBS Lett. 400, 103–107.PubMedCrossRefGoogle Scholar
  8. 8.
    Belozersky, M. A., Dunaevsky, Y. E., Musolyamov, A. V., and Egorov, T. A. (1995), FEBS Lett. 371, 264–266.PubMedCrossRefGoogle Scholar
  9. 9.
    Kouzuma, Y., Suetake, M., Kimura, M., and Yamasaki, N. (1992), Biosci. Biotechnol. Biochem. 56, 1819–1824.PubMedCrossRefGoogle Scholar
  10. 10.
    Sallay, I., Moriwaki, S., Nakamura, O., Yasuda, S., Kimura, M., Yamasaki, N., Itoh, K., and Ohba, H. (2000), J. Hematother. Stem Cell Res. 9, 877–883.PubMedCrossRefGoogle Scholar
  11. 11.
    Mosmann, T. (1983), J. Immunol. Methods 65, 55–63.PubMedCrossRefGoogle Scholar
  12. 12.
    Patty, L. and Smith, E. L. (1975), J. Biol. Chem. 250, 557–564.Google Scholar
  13. 13.
    Ikeda, K. and Kusano, T. (1978), Agric. Biol. Chem. 42, 309–314.Google Scholar
  14. 14.
    Shibata, H., Hara, S., and Ikenaka, T. (1986), J. Biochem. 99, 1147–1155.PubMedGoogle Scholar
  15. 15.
    Stanley, P. (1981), in The Biochemistry of Glycoproteins and Proteoglycans, Lennarz, W. J., ed., Plenum, New York, pp. 161–190.Google Scholar
  16. 16.
    Kim, M., Rao, M. V., Tweardy, D. J., Prakash, M., Galili, U., and Gorelik, E. (1993), Glycobiology 3, 447–453.PubMedCrossRefGoogle Scholar
  17. 17.
    Moriwaki, S., Ohba, H., Nakamura, O., Sallay, I., Suzuki, M., Tsubouchi, H., Yamasaki, N., and Itoh, K. (2000), J. Hematother. Stem Cell Res. 9(1), 47–53.PubMedCrossRefGoogle Scholar
  18. 18.
    Yavelow, J., Caggana, M., and Beck, K. A. (1987), Cancer Res. 47, 1598–1601.PubMedGoogle Scholar
  19. 19.
    Yavelow, J., Scott, C. B., and Mayer, T. C. (1987), Cancer Res. 47, 1602–1607.PubMedGoogle Scholar

Copyright information

© Humana Press Inc. 2004

Authors and Affiliations

  1. 1.Cheju Traditional Food InstituteCheju Halla CollegeJeju-doKorea
  2. 2.Natural Substance-Composed Material GroupNational Institute of Advanced Industrial Science and TechnologySagaJapan

Personalised recommendations