Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Effects of Aralia continentalis Root Extract on Cell Proliferation and Apoptosis in Human Promyelocytic Leukemia HL-60 Cells

  • Lim Hae-Young (Department of Medical Genetics, College of Medicine, Hanyang University) ;
  • Oh Ha-Lim (Department of Medical Genetics, College of Medicine, Hanyang University) ;
  • Lee Chul-Hoon (Department of Medical Genetics, College of Medicine, Hanyang University)
  • Published : 2006.09.01
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Abstract

The roots of Aralia continentalis (AC) have been used traditionally in Korean as a folk medicine for anti-inflammation and as an anti-rheumatic. In this study, we report that the ethyl acetate-soluble traction (ACE) of the methanolic extract of AC root inhibited the cell growth of various human cancer cell lines and induced apoptosis of HL-60, human promyelocytic leukemia cells. Its $IC_{50}$ values on growth inhibition were estimated to be $56.3{\mu}g/ml$ on HL-60, $87.2{\mu}g/ml$ on HepG2, $93.2{\mu}g/ml$ on HeLa, $135.5{\mu}g/ml$ on DU-145, and $135.8{\mu}g/ml$ on HT-29 cells. Interestingly, ACE showed no antiproliferative effect on normal lymphocyte cells used as control. Furthermore, nuclear DAPI staining revealed the typical nuclear features of apoptosis in the HL-60 cells exposed to $80{\mu}g/ml$ ACE, and a flow cytometric analysis of the HL-60 cells using propidium iodide showed that the apoptotic cell population increased gradually from 5% at 0 h to 16% at 12 h and 20% at 24 h after treated with $50{\mu}g/ml$ of ACE. TUNEL assay also revealed the apoptotic induction of the HL-60 cells treated with ACE. To obtain further information on the ACE-induced apoptosis, the expression level of certain apoptosis-associated proteins was examined using a Western blot analysis. Treatment of the HL-60 cells with ACE resulted in the activation of caspase-3, and subsequent proteolytic cleavage of poly(ADP-ribose) polymerase (PARP). The above results confirmed that the apoptosis in the HL-60 cells was induced by ACE, and that caspase-3-mediated PARP cleavage was involved in the process.

Keywords

References

  1. Brown, J. M. and B. G. Wouters. 1999. Apoptosis, p53, and tumor cell sensitivity to anticancer agents. Cancer Res. 59: 1391-1399
  2. Coleman, M. L., E. A. Sahai, M. Yeo, M. Bosch, A. Dewar, and M. F. Olson. 2001. Membrane blebbing during apoptosis results from caspase-mediated activation of ROCK I. Nat. Cell Biol. 3: 339-345 https://doi.org/10.1038/35070009
  3. Dantzer, R., V. Schreiber, C. Niedergang, C. Trucco, E. Flatter, G De La Rubia, J. Oliver, V. Rolli, J. Menissier-de Murcia, and G. de Murcia. 1999. Involvement of poly(ADP-ribose) polymerase in base excision repair. Biochimie 81: 69-75 https://doi.org/10.1016/S0300-9084(99)80040-6
  4. Enari, M., H. Sakahira, H. Yokoyama, K. Okawa, A. Iwamatsu, and S. Nagata. 1998. A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature 391: 43-50 https://doi.org/10.1038/34112
  5. Han, B. H., Y. N. Han, K. A. Han, M. H. Park, and E. O. Lee. 1983. Studies on the anti-inflammatory activity of Aralia continentalis (I). Arch. Pharm. Res. 6: 17-23 https://doi.org/10.1007/BF02855697
  6. Han, B. H., E. R. Woo, M. H. Park, and Y. N. Han. 1985. Studies on the anti-inflammatory activity of Aralia continentalis (III). Arch. Pharm. Res. 8: 59-65 https://doi.org/10.1007/BF02912223
  7. Hengartner, M. O. 2000. The biochemistry of apoptosis. Nature 407: 770-776 https://doi.org/10.1038/35037710
  8. Kaufman, S. H. 1989. Induction of endonucleolytic DNA cleavage in human acute myelogenous leukemia cells by etoposide, camptothecin and other cytotoxic anticancer drugs: A cautionary note. Cancer Res. 49: 5870-5878
  9. Kaufman, S. H. and W. C. Earnshaw. 2000. Induction of apoptosis by cancer chemotherapy. Exp. Cell Res. 256: 42-49 https://doi.org/10.1006/excr.2000.4838
  10. Kerr, J. F. R., A. H. Wyllie, and A. R. Currie. 1972. Apoptosis: A basic biological phenomenon with wide-ranging implication in tissue kinetics. Brit. J. Cancer 26: 239-257 https://doi.org/10.1038/bjc.1972.33
  11. Kim, M., C. Shin, H. Yang, S. Kim, H. Lim, C.-H. Lee, M. Kim, and Y Lim. 2004. Naltriben analogues as peptide anticancer drugs. J. Microbiol. Biotechnol. 14: 881-884
  12. Lee, C.-H., H. Lim, M. Kim, Y. Cho, D. Oh, C. Kim, and Y. Lim. 2002. Isolation and biological properties of novel cell cycle inhibitor, HY558, isolated from Penicillium minioluteum F558. J. Microbiol. Biotechnol. 12: 470-475
  13. Lee, H.-K., I.-H. Lee, J.-S. Yim, Y.-H. Kim, S.-H. Lee, K.-S. Lee, Y.-M. Koo, S.-J. Kim, and B.-C. Jeong. 2005. Purification and characterization of CDMHK, a growth inhibitory molecule against cancer cell lines, from Myxobacterium sp. HK1 isolated from Korean soil. J. Microbiol. Biotechnol. 15: 734-739
  14. Lim, H., M. Kim, Y. Cho, J. Kim, Y. Lim, and C.-H. Lee. 2004. Inhibition of cell cycle progression and induction of apoptosis in HeLa cells by HY558-1, a novel CDK inhibitor isolated from Penicillium minioluteum F558. J. Microbiol. Biotechnol. 14: 978-984
  15. Lim, H., M. Kim, Y. Lim, Y. Cho, and C.-H. Lee. 2006. Inhibition of cell-cycle progression in HeLa cells by HY52, a novel cyclin-dependent kinase inhibitor isolated from Bauhiniaforficata. Cancer Lett. 233: 89-97 https://doi.org/10.1016/j.canlet.2005.03.002
  16. Lim, T. Y 1987. Comparative study on the pharmacological effect of kinds of Dokhwal. J. Kor. Orient. Med. 8: 97-98
  17. Los, M., C. J. Burek, C. Strol, K. Bnedyk, H. Hug, and A. Mackiewicz. 2003. Anticancer drugs of tomorrow: Apoptotic pathways as targets for drug design. Drug Discov. Today 8: 67-77 https://doi.org/10.1016/S1359-6446(02)02563-1
  18. Mosmann, T. 1983. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J. Immunol. Methods 65: 55-63 https://doi.org/10.1016/0022-1759(83)90303-4
  19. Nunez, G, M. A. Benedict, Y. Hu, and N. Inohara. 1998. Caspases: The proteases of the apoptotic pathway. Oncogene 17: 3237-3245 https://doi.org/10.1038/sj.onc.1202581
  20. Reed, J. C. 2001. Apoptosis regulatory proteins as targets for drug discovery. Trends Mol. Med. 7: 314-318 https://doi.org/10.1016/S1471-4914(01)02026-3
  21. Sahara, S., M. Aoto, Y. Eguchi, N. Imamoto, Y. Yoneda, and Y. Tsujimoto. 1999. Acinus is a caspase-3-activated protein required for apoptotic chromatin condensation. Nature 401: 168-173 https://doi.org/10.1038/43678
  22. Sebbagh, M., C. Renvoize, J. Hamelin, N. Riche, J. Bertoglio, and J. Breard. 2001. Caspase-3-mediated cleavage of ROCK I induces MLC phosphorylation and apoptotic membrane blebbing. Nat. Cell Biol. 3: 346-352 https://doi.org/10.1038/35070019
  23. Suh, N., L. Luyengi, H. H. Fong, A. D. Kinghorn, and J. M. Pezzuto. 1995. Discovery of natural product chemopreventive agents utilizing HL-60 cell differentiation as a model. Anticancer Res. 15: 233-239
  24. Studzinski, G P. 1999. An overview of apoptosis. In G P. Studzinski (ed.), Apoptosis: A Practical Approach. Oxford University Press, Oxford, U.K