Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Induction of Apoptotic Cell Death in Human Jurkat T Cells by a Chlorophyll Derivative (Cp-D) Isolated from Actinidia arguta Planchon

  • Park, Youn-Hee (Department of Microbiology, College of Natural Science, Kyungpook National University) ;
  • Chun, En-Mi (Department of Microbiology, College of Natural Science, Kyungpook National University) ;
  • Bae, Myung-Ae (Department of Microbiology, College of Natural Science, Kyungpook National University) ;
  • Seu, Young-Bae (Department of Microbiology, College of Natural Science, Kyungpook National University) ;
  • Song, Kyung-Sik (Department of Agricultural Chemistry, College of Agriculture, Kyungpook National University) ;
  • Kim, Young-Ho (Department of Microbiology, College of Natural Science, Kyungpook National University)
  • Published : 2000.02.01
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Abstract

The chloroform and methanol (2;1, v/v) extract from an edible plant, Actinidia arguta Planchon, appeared to possess antitumor activity against human leukemias Jurkat T and U937 cells through inducing apoptosis. The substance in the solvent extract was purified by silica gel column chromatography, preparative TLC, and Sephadex LH-20 column chromatography. Characteristics of the substance analyzed by UV scanning analysis, $^1H$ and $^{13}C$ NMR spectra suggested that the substance belongs to the chlorophyll derivatives-like group. The $IC_{50}$ value of the chlorophyll derivative (Cp-D) determined by MTT assay was $15\mu\textrm{g}/ml$ for Jurkat, $10\mu\textrm{g}/ml$ for U937, and $11.4\mu\textrm{g}/ml$ for HL-60m and was more toxic to these leukemias than to solid tumors or normal fibroblast. In order to elucidate cellular mechanisms underlying the cytotoxicity, the effect of the Cp-D on Jurkat T cells was investigated. When cells were treated with the Cp-D at a concentration of $15\mu\textrm{g}/ml$, [3H]thymidine incorporation declined rapidly and wa undetectable in 1h. However, no significant changes were made in the cell cycle distribution of the cells by 24h. The sub-Gl peak representing apoptotic cells began to be detectable in 36h, at which time apoptotic DNA fragmentation was also detected on agarose gel electrophoresis, demonstrating that the cytotoxic effect of the Cp-D is attributable to the induced apoptosis. Under the same conditions, although the protein level of cyclin-dependent kinases such as cdc4, csk6, cdk2, and cdc2 was not significantly changed until 24h, the kinase activity of all c안 rapidly declined and reached a minimum level within 1-6h and then recovered to the initial level by 12h and sustained until 24h. These results suggest that inactivation of cdks at an inappropriate time during the cell cycle progression in jurkat T cells following a treatment with the Cp-D leads to induction of apoptotic cell death.

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References

  1. Biochem. Biophys. Res. Commun. v.246 Enhanced expression of Fas ligand is associated with aburatubolactam C-induced apoptosis in human Jurkat T cell. Bae, M. A.;H. S. Kang;S. W. Rue;J. H. Seu;Y. H. Kim
  2. J. Microbiol. Biotechnol. v.8 Aburatubolactam C, a novel apoptosis inducing substance produced by marine Streptomyces sp. SCRC-20 Bae, M. A.;K. Yamada;D. Uemura;J. H. Seu;Y. H. Kim
  3. Cell v.78 Apoptosis in cancer therapy: Crossing the threshold Fisher, D. E.
  4. Oncogene v.11 Cell cycle control in mammalian cells: Role of cyclins, cyclin dependent kinases(CDKs), growht suppressor genes and cyclin-dependent kinase inhibitors (CKIs). Grana, X;E. P. Reddy
  5. Blood v.89 Apoptosis and dilemma of cancer chemotherapy. Hannun, Y. A.
  6. Obstet. Gynecol. v.85 Chemotherapy-induced apoptosis in epithelial ovarian cancers. Havrilesky, L. J.;A. Elbendary, J. A.;Hurteau;R. S. Whitaker;G.C. Rodriguez;A. Berchuck.
  7. Nucleic Acids Res. v.22 A rapid and simple method for the isolation of apoptotic DNA fragments. Herrmann, M;H.0M. Lorenz;R. Voll;M. Granke;W. Woith;J. R. Kalden
  8. Cell v.79 Cyclins and cancer; cyclin D and cdk inhibitors come of age. Hunter, T.;J. Pines.
  9. Br. J. Cancer v.73 Characteristics of cancer cell death after exposure to cytotoxic drugs in vitro Hushtscha, L. I.;W. A. Bartier, C. E. Ross.;M. H. Tattersall.
  10. Mol. Cells v.8 Characterization of the murine cdc2 gene. Jun, D. Y.;H. K. Park;A. A. Nordin;J. E. Nagel;Y. H. Kim
  11. Cancer Res v.49 Induction of endonucleolytic DNA cleavage in human acute myelogenous leukemia cells by etoposide, camptothecin, and other cytotoxic DNA cleavage in human acute myelogenous leukemia cells by etoposide, camptothecin, and other cytotoxic anticancer drugs: A cautionary note Kaufman, S. H.
  12. J. Immunol. v.158 Expression of the murine homologue of the cell cycle control protein $p34^cdc2$ in T lymphocytes. Kim, Y. H.;J.J. Proust;M.J. Buchholz;F. J. Chrestr;A. A. Nordin
  13. J. Immunol. v.158 The regulation of $P^Kip1$ expression following the polyclonal activation of murine $G_0T$ cells Kwon, T. K.;M. A. Buchholz;P. Ponsalle;F. J. Chrest;A. A. Nordin
  14. Clinical Cancer Medicine:Treatment Tactics Lokich, J. J.
  15. Magn. Reson. Chem. v.25 High-field proton NMR study of the liquid solution structure of monomeric bacteriochlorophyll a and chlorophyll a Lotjonen, S.;T. J. Michalski;J.R. Norris;P. H. Hynninen
  16. Proc. Natl. Acad. Sci. v.91 Activation of cyclin A-dependent protein kinases during apoptosis. Meikrants, W.;S. Gisselbrecht;S. W. Tam;R. Schiegel.
  17. Cell v.79 Joining the complex: cyclin depedent kinase inhibitoty proteins and the cell cycle. Peter, M.;I. Herskowltz.
  18. J. Immunol. v.157 Granzyme B induces apoptosis and cyclin A-associated cyclin-dependent kinase activity in all stages of the cell cycle Shi, L;G. Chen;D. He;D. G. Bosc;D.W. Litchifield;A. H. Greenberg
  19. Cell v.79 G1 phase progression Cycling on cue Sherr, C. J.
  20. Science v.263 Premature $p34^cdc2$ activation required for apoptosis. Shi, L.;W. K. Nishioka;J. M. Th'ng;E. M. Bradbary;D.W. Litchfield;A. H. Greenberg
  21. Exp. Cell. Res. v.210 Enhancement of CDDP cytotoxicity by caffeine is characterized by apoptotic cell death. Shinomiya, N;M. Shinomiya;H. Wakiyama;Y. Katsura;M. Rokitanda
  22. Org. Mag. Reson. v.21 no.` $Carbon-^-13NMR$ spectra of chlorophyll a chlorophyll a', phrochlorophyll a and the corresponding pheophytins. Simo, L;P. H. Hynninen
  23. Cancer Res. v.51 Topoisomerase Ⅱ-reactive cchemotherapeutic drugs induce apoptosis in thymocytes. Walker, P. R.;C. Smith;T. Youdale;J. Lebnaris;J. F. Whitfield;M. Sikorska
  24. Cell Growth Differ v.6 Apoptosis in 7-hydroxystaurospurine-treated T lymphoblasts correlates with activation of cyclin dependent kinases 1 and 2 Wang, Q;P.J. Worland;J. L. Clark;B. A. Carlson;E. A. Sausbille
  25. J. Biol. Chem. v.271 The cooperative interaction of two different signalling pathways in response to byfalin induces apoptosis in human leukemia U937 cell. Watabe, M. Y.;Mansuda;S. Nakajo;T. Yoshida;T. Kuroiwa;K. Nakaya
  26. Cell v.65 Programmed cell death: Apoptosis and Oncogenesis Williams, G.T.
  27. Methods in Natural Products Chemistry(1st ed) Woo, W.-S.
  28. Cancer Res. v.56 Requirement of Yao, S. L.;K. A. Mekenna;S. J. Sharkis;A. Bedi $p34^cdc2$ kinase for apoptosis mediated by the Fas/Apo-1 receptor and interleukin 1β-converting enzyme related proteases.
  29. Dev. Biol. v.183 Cyclin depedent kinase 5 is associated with apoptotic cell death during development and tissue remodelling. Zhang, Q;H. S. Ahuka;Z. F. Zakeri