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Growth Inhibitory and Quinone Reductase Activity Stimulating Effects of Internal Organs of Aplysia kurodai Fractions on Cancer Cell Lines In vitro

군소내장 분획물의 in vitro에서의 암세포 성장억제 및 quinone reductase 유도 활성 증가 효과

  • Shin, Mi-Ok (Dept. of Food and Nutrition, Silla University)
  • 신미옥 (신라대학교 식품영양학과)
  • Received : 2010.02.17
  • Accepted : 2010.03.30
  • Published : 2010.06.30

Abstract

We investigated the growth inhibitory effect of internal organs of Aplysia kurodai (AK) on proliferation in cancer cell lines in vitro. The internal organs of AK were extracted with methanol (AKM), which were then further fractionated into four subfractions by using solvent partition method, resulting in hexane (AKMH), methanol (AKMM), butanol (AKMB), and aqueous (AKMA) soluble fractions. We determined the cytotoxic effect of these four fractions in four kinds of cancer cell lines - HepG2, MCF-7, HT29 and B16-F10 - by MTT assay. Among the four subfractions of AKM, AKMM showed the strongest cytotoxic effects on all cancer cell lines which were used. Morphological changes such as membrane shrinking and blebbing of cells were also observed in AKMM treatment in HepG2 cells. In addition, we also observed quinone reductase (QR) induced effect in the methanol layer (AKMM) of HepG2 cells. AKMM showed the highest induction activity of quinone reductase on HepG2 cells among the partition layers. The QR induced effect of AKMM was determined to be 2.4 at $100\;{\mu}g/ml$ level with a control value of 1.0. Although further studies are needed, the present work suggests that internal organs of Aplysia kurodai (AK) may be a chemopreventive agent for the treatment of human cells.

본 연구에서는 군소내장을 각 용매별로 분획하여 암세포 성장 억제 효과와 quinone reductase (QR) 유도 활성증가 효과를 알아보았다. 간암 세포인 HepG2, 유방암 세포주인 MCF-7, 대장암 세포주인 HT-29 그리고 피부암 세포주인 B16-F10 를 이용하여 암세포 성장 억제 효과를 실험한 결과 모든 세포주에서 AKMM층에서 농도 의존적으로 가장 높은 암세포 성장 억제 효과를 나타내었다. 그리고 다음으로 AKMB층, AKMH층 및 AKMA층의 순이었다. 그리고 4종의 암세포주 중에서 B16-F10 세포주가 가장 높은 암세포 성장 억제 효과를 나타내어 특히 피부암에 대한 예방효과가 기대되어진다. 또한 암 예방 효과를 알아보기 위하여 4종의 암세포 중 유일하게 quinone reductase를 가지고 있는 인체 간암세포주인 HepG2를 이용하여 QR 유도 활성 증가 효과를 측정한 결과, 암세포 성장 억제 효과에 있어서 가장 높은 효과를 나타낸 AKMM층에서 가장 높은 QR 유도 활성 증가 효과를 나타내었다. 그러나 다른 분획층에서는 QR 유도 활성 증가 효과를 거의 볼 수 없었다. 이상으로 암세포 성장 억제 효과와 QR 유도 활성 효과에서 모두 methanol 분획층인 AKMM층에서 가장 높게 나타났으므로 이 분획층에 유효한 생리활성 물질이 함유되어 있을 가능성이 추정되어진다. 따라서 폐기되어지는 군소부산물인 내장을 이용한 암 예방 관련 기능성 식품의 개발 가능성이 기대되어지며, 이를 위한 AKMM분획층에 대한 더욱더 심도 높은 집중적인 연구가 요구되어진다.

Keywords

References

  1. Alley, M. C., D. A. Scudiero, A. Monks, M. L. Hursey, M. J. Czerwinski, D. L. Fine, B. J. Abbortt, J. G. Mayo, R. H. Shoemarker, and M. R. Boyd. 1988. Feasibility of drug screening with panels of human tumor cell lline using a microculture tetrazolium assay. Cancer Res. 48, 589-601.
  2. Bae, M. J., S. T. Yee, S. Y. Chae, S. H. Shin, S. H. Kweon, M. H. Park, M. K. Song, and S. J. Hwang. 2004. The effect of arabinoxylane and the polysaccharide peptide (PSP) on the antiallergy, anticancer. J. Korean Soc. Food Sci. Nutr. 33, 469-474. https://doi.org/10.3746/jkfn.2004.33.3.469
  3. Banner, S. E., U. Pastorino, S. M. Lippman, and W. K. Hong. 1994. Second international chemoprevention conference. Cancer Res. 54, 854-860.
  4. Budd, G. T., B. Osgood, B. Barna, I. M. Boyett, J. Finke, S. V. Mdendrop, S. Murth, and R. M. Bukoski. 1989. Phase I clinical trial Toxicity and immunologic effects, Cancer Res. 49, 6432-6439.
  5. Carmichael, J., W. G. De Graff, A. F. Gazder, J. D. Minna, and J. B. Mitchell. 1987. Evalution of the tetrazolium based semiautomated colorimetric assay: assessment of chemosensitivity testing. Cancer Res. 47, 936-942.
  6. Cha, B. C., H. W. Lee, and M. Y. Cho. 1998. Antioxidative and antimicrobial effeats of nut species. Kor. J. Pharmacogn. 29, 28-34.
  7. Choe, B. L. and J. R. Lee. 1994. Opisthobranchs (mollusca: gastropoda) from ullung and dog-do islands, korea. Korean J. Zool. 37, 352-376.7.
  8. Giampietri, A. 1981. Drug-mediated increase of tumor immunogenicity in vivo for a new approach to experimental cancer immunotherapy. Cancer Res. 41, 681-685.
  9. Giampietri, A. 1981. Drug-mediated increase of tumor immunogenicity in vivo for a new approach to experimental cancer immunotherapy. Cancer Res. 41, 681-685.
  10. Hong, E. Y., H. J. Kang, C. S. Kwan, Y. J. Na, M. J. Suh, and J. S. Kim. 1997. Modulation of cellular quinone reductase inducibility by roasting treatment and acid hydrolysis of perilla. J. Korean Soc. Food Sci. Nutr. 26, 186-192.
  11. Jung, Y. H., B. M. Jung, M. O. Shin, and S. J. Bae. 2006. A study on the effects of anticarcinogenic activity of gloiopeltis tenax. J. Korean Soc. Food Sci. Nutr. 35, 395-401. https://doi.org/10.3746/jkfn.2006.35.4.395
  12. Kandel, E. R., J. H. Schwartz, and T. M. Jessell. 2000. Principles of neural science, 4th eds. McGraw-Hill, New York, 180.
  13. Kim, C. H., H. J. Seo, E. Y. Hwang, E. J. Kim, H. J. Go, I. H. Kim, J. K. Seo, J. H. Moon, M. D. Huh, and N. G. Park. 2001. Purification of myomoduline A and myomodulin E from the central nervous system of the sea lare, Aplysia kurodai. J. Korean Fish Soc. 34, 279-284.
  14. Kim, H. S. and B. L. Choe. 1981. The founa of marine invertebrate in ulreung is. and dog-do is. special report. The Conservation of Nature and Natural Resources 19, 193-200.
  15. Kim, J. S. 2005. Cancer chemoprevention and NSAID-activated gene (NAG-1). Biochemistry and Molecular Biology News 12, 286-291.
  16. Lee, J. S. and D. K. Min. 2002. A catalogue of molluscan fauna in Korea. Opisthobranchs 18, 93-97.
  17. Lee, Y. S., D. S. Kim, B. H. Ryu, and S. H. Lee. 1992. Antitumor and Immunodulating effect of seaweeds toward sarcoma-180 cell. J. Korean Soc. Food Sci. Nutr. 21, 544-550.
  18. Lloyd, P. E., I. Kuufermann, and K. R. Weiss. 1987. Sequence of small cardioactive peptide A: A second member of a class of neuropeptides in Aplysia. Peptides 8, 179-184. https://doi.org/10.1016/0196-9781(87)90184-7
  19. Morris, H. R., M. Panico, A. Karplus, P. E. Lloyd, and B. Riniker. 1982. Elucidation by FAB-MS of the structure of a new cardioactive peptide from Aplysia. Nature 300, 643-645. https://doi.org/10.1038/300643a0
  20. Nishiwaki, S., H. Ueda, and T. Makioka. 1975. Tagging studies on the growth of the sea hare Aplysia kurodai on an intertidal rocky shore. Marine Biology 32, 389-395. https://doi.org/10.1007/BF00388996
  21. Park, H. J. 1998. Induction of quinone reductase and its regulatory mechanism at the transcriptional level by Scutellaria baicalensis, Ph. D. Dissertation, Yonsei University, Seoul.
  22. Park, J. C. and J. W. Choi. 1996. Screening of marine natural products on inhibitory effect of the formation of lipid peroxidation. Kor. J. Pharmacogn. 27, 117-122.
  23. Park, S. Y., B. M. Jung, Y. H. Choi, and S. J. Bae. 2005. Growth inhibition effects of cancer cell lines by gloiopeltis furcata fractions in vitro. J. Korean Soc. Food Sci. Nutr. 34, 771-775. https://doi.org/10.3746/jkfn.2005.34.6.771
  24. Park, Y. J., M. O. Shin, S. H. Lee, and S. J. Bae. 2005. The growth inhibitory effects of atrina pecitinata fractions on cancer cell lines. Korean J. Nutrition 38, 307-312.
  25. Prochaska, H. J. 1994. Screening strategies for the detection of anticarcinogenic enzyme inducers. J. Mutr. Biochem. 5, 360-370. https://doi.org/10.1016/0955-2863(94)90067-1
  26. Prochaska, H. J., A. B. Santamaria, and P. Talalay. 1992. Rapid detection of inducers of enzyme that protect against carcinogene. Proc. Natl. Acad. Sci. USA 89, 2394-2399. https://doi.org/10.1073/pnas.89.6.2394
  27. Prochaska, H. J. and A. B. Santamaria. 1988. Direct measurement of NAD(P)H : Quinone reductase from cells cultured in microtiter wells : A screening assay for anticarcinogenic enzyme inducers. Anal. Biochem. 169, 328-336. https://doi.org/10.1016/0003-2697(88)90292-8
  28. Schwartsmann, G., A. B. Roch, R. G. Berlinck, and J. Jimeno. 2001. Marine organisms as a source of new anticancer agents. Oncology 2, 221-225. https://doi.org/10.1159/000214620
  29. Shin, M. O., M. J. Ku, and S. J. Bae. 2007. Cytotoxicity and quinone reductase activity stimulating effects of fin of thunnus thynnus extracts in various cancer cells. Korean J. Nutrition 40, 147-153.
  30. Shin, M. O. and S. J. Bae. 2009. Growth inhibitory and quinone reductase activity stimulating effects of internal organs of todarodes pacificus fractions on human cancer cell lines in vitro. J. Life Science 19, 1251-1257. https://doi.org/10.5352/JLS.2009.19.9.1251
  31. Shin, M O. and S. J. Bae. 2009. The anticarcinogenic and antioxidative activity of Hemicentrotus pulacherrimus fractions in various cancer cells. J. Life Science 19, 607-614. https://doi.org/10.5352/JLS.2009.19.5.607
  32. Shon, J. H., D. Y. Kang, H. C. OH, B. M. Jung, M. H. Kim, M. O. Shin, and S. J. Bae. 2006. The effects on antimicrobal and cytotoxicity of hijikia fusiformis fraction. J. Korean Soc. Food Sci. Nutr. 39, 444-450.
  33. Shon, Y. H., K. H. Jeune, and S. J. Choi. 1998. Chung S. R. Life Science; Antitumor effect of asterina pecinifera lectin on ascitic tumor. Yakhak Hoeji 42, 99-105.
  34. Tusa, Y. 1994. Size-related egg production in a simultaneous hermaphrodite, the sea hare Aplysia kurodai baba (mollusca: opisthobranchia). Publ. Seto Mar. Biol. 36, 249-254.
  35. Wefers, H., T. Komai, P. Talalay, and H. Sies. 1984. Protection against reactive oxygen species by NAD(P)H: quinone reductase induced by the dietary antioxidant butylated hydroxyanisole (BHA). Federation of European Biochemical Societies 169, 63-66. https://doi.org/10.1016/0014-5793(84)80290-2