Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
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Effect of the Hexane Extract of Saussurea lappa on the Growth of HT-29 Human Colon Cancer Cells

목향 헥산추출물이 대장암세포인 HT-29 세포의 증식에 미치는 영향

  • Kim, Eun-Ji (Center for Efficacy Assessment and Development of Functional Foods and Drugs, Hallym University) ;
  • Park, Hee-Sook (Department of Food Science and Nutrition) ;
  • Lim, Soon-Sung (Center for Efficacy Assessment and Development of Functional Foods and Drugs, Hallym University) ;
  • Kim, Jong-Sang (Department of Animal Science and Biotechnology, Kyungpook National University) ;
  • Shin, Hyun-Kyung (Center for Efficacy Assessment and Development of Functional Foods and Drugs, Hallym University) ;
  • Yoon, Jung-Han (Center for Efficacy Assessment and Development of Functional Foods and Drugs, Hallym University)
  • 김은지 (한림대학교 식의약품의 효능평가 및 기능성소재개발센터) ;
  • 박희숙 (한림대학교 식품영양학과) ;
  • 임순성 (한림대학교 식의약품의 효능평가 및 기능성소재개발센터) ;
  • 김정상 (경북대학교 동물공학과) ;
  • 신현경 (한림대학교 식의약품의 효능평가 및 기능성소재개발센터) ;
  • 윤정한 (한림대학교 식의약품의 효능평가 및 기능성소재개발센터)
  • Published : 2008.04.30
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Abstract

In Asia Saussurea lappa (SL) has been used as a traditional herbal medicine to treat abdominal pain and tenesmus. Recently, in vitro cell culture studies have shown that SL has anti-ulcer, anti-inflammatory, and anti-tumor properties. To explore its potential chemopreventive and chemotherapeutic effects in colon cancer, we examined whether the hexane extract of SL (HESL) could inhibit the growth of HT-29 human colon cancer cells, and investigated the mechanisms for this effect. The cells were cultured with various concentrations (0-5 ${\mu}g/mL$) of HESL. The results indicated that HESL markedly decreased the numbers of viable HT-29 cells; whereas at the concentration of 5 ${\mu}g/mL$, HESL slightly decreased the viable cell numbers of CCD 1108Sk human skin normal fibroblasts at 72 hr. HESL substantially increased the numbers of cells in the sub G1 phase, and dose-dependently increased apoptotic cell numbers. Western blot analysis of the total cell lysates revealed that HESL increased Bax protein levels, but did not affect Bcl-2 levels. HESL induced the cleavage of poly (ADP-ribose) polymerase and caspases 8, 9, 7, and 3. This study demonstrated that HESL inhibits cell growth and induces apoptosis in HT-29 cells, which may be mediated by its ability to increase Bax levels and activate the caspase pathway. These findings may lead to the development of new therapeutic strategies for colon cancer treatment.

목향(Saussurea lappa)은 항암효과를 비롯하여 다양한 생리활성을 나타내는 것으로 알려지고 있으나 목향이 대장암에 미치는 영향에 대해 자세히 연구된 바가 없다. 본 연구에는 목향 헥산추출물이 인간의 대장에서 유래한 암세포인 HT-29 세포의 증식에 미치는 영향과 그 작용 기전에 대해 연구하였다. 헥산으로 목향을 추출하여 얻은 목향 헥산추출물을 HT-29 세포의 세포 배양액에 0, 1, 3, 5 ${\mu}g/mL$로 첨가하여 세포를 배양하였다. HT-29 세포의 증식은 목향 헥산추출물 처리 농도가 증가할수록 현저히 감소하였다. 인간의 대장에서 유래한 정상 상피세포인 FHC 세포의 증식은 목향 헥산추출물에 의해 변화하지 않았고, 인간의 피부에서 유래한 정상 섬유아세포인 CCD 1108Sk 세포 증식은 목향을 5${\mu}g/mL$ 농도로 72 시간 배양한 경우에만 감소하였다. 목향 헥산 추출물 처리에 의해 HT-29 세포의 세포주기 진행 중 sub G1기에 머무른 세포수가 증가하였고, 세포사멸 세포수가 현저히 증가하였다. 세포사멸의 주요한 조절인자인 Bcl-2 family 단백질 중 Bcl-2은 목향 헥산추출물에 의해 변화하지 않았으나 Bax는 유의적으로 증가하였다. Bcl-2 family 단백질과 더불어 세포사멸 조절에 중요한 역할을 하는 caspases의 활성형인 cleaved caspase-8, -9, -7, -3가 목향 헥산추출물에 의해 증가하였다. 또한 목향 헥산추출물 처리 농도가 증가할수록 cleaved PARP 단백질 수준도 현저히 증가하였다. 이 결과는 목향 헥산추출물이 세포사멸을 유도하여 대장암세포인 HT-29 세포의 증식을 억제함을 나타내며, 목향 헥산추출물에 의해 HT-29 세포의 세포사멸은 Bax 증가와caspases의 활성 증가에 의한 것임을 나타낸다. 본 연구는 목향을 독성과 부작용이 적은 암예방제나 항암제로 개발할 수 있는 가능성을 제시한다. 그러나 목향을 이용하여 제품 개발을 위해서는 유효 성분 동정 및 동물시험 등의 연구 수행이 필요할 것으로 사료된다.

Keywords

References

  1. Jemal A, Siegel R, Ward E, Murray T, Xu J, Thun MJ. Cancer statistics, 2007. CA Cancer J. Clin. 57: 43-66 (2007) https://doi.org/10.3322/canjclin.57.1.43
  2. Lee KH, Yu CS, Kim HC, Kim JR, Kim YM, Kim JS, Kim JC. Outcome of curative resection in patients with completely obstructing colorectal cancer. J. Korean Surg. Soc. 66: 199-204 (2004)
  3. Kim WS, Lee RA, Hwang DY, Hong YJ, Hong SI. Histoculture drug response assay in colorectal cancer specimen. J. Korean Surg. Soc. 66: 109-115 (2004)
  4. Maroun JA, Anthony LB, Blais N, Burkes R, Dowden SD, Dranitsaris G, Samson B, Shah A, Thirlwell MP, Vincent MD, Wong R. Prevention and management of chemotherapy-induced diarrhea in patients with colorectal cancer: A consensus statement by the canadian working group on chemotherapy-induced diarrhea. Curr. Oncol. 14(1): 13-20 (2007) C https://doi.org/10.3747/co.2007.96
  5. de Gramount A, Figer A, Seymour M, Homerin M, Hmissi A, Cassidy J, Boni C, Cortes-Funes H, Cervantes A, Freyer G, Papamichael D, Le Bail N, Louvet C, Hendler D, de Braud F, Wilson C, Morvan F, Bonetti A. Leucovorin and fluorouracil with or without oxaliplatin as first-line treatmet in advanced colorectal cancer. J. Clin. Oncol. 18: 2938-2947 (2000) https://doi.org/10.1200/JCO.2000.18.16.2938
  6. Jeon YJ, Lee HS, Yeon SW, Ko JH, An KM, Yu SW, Kang JH, Hwang BY, Kim TY. Inhibitory effects of dehydrocostuslactone isolated from Saussureae Radix on CDK2 activity. Korean J. Pharmacogn. 36(2): 97-101 (2005)
  7. Jung JH, Ha JY, Min KR, Shibata F, Nakagawa H, Kang SS, Chang IM, Kim YS. Reynosin from Saussurea lappa as inhibitor on CINC-1 induction in LPS-stimulated NRK-52E cells. Planta Med. 64: 454-456 (1998) https://doi.org/10.1055/s-2006-957480
  8. Jung JH, Kim YS, Lee CO, Kang SS, Park JH, Im KS. Cytotoxic constituents of Saussurea lappa. Arch. Pharm. Res. 21: 153-156 (1998) https://doi.org/10.1007/BF02974020
  9. Kang SS, Kim JS, Chi HJ, Chang SY, Ha KW. Isolation and quantitative determination of costunolide from Saussurea root. Korean J. Pharmacogn. 30: 48-53 (1999)
  10. Yoshikawa M, Hatakeyama S, Inoue Y, Yamahara J. Saussureamines A, B, C, D, and E, new anti-ulcer principles from Chinese Saussureae. Chem. Pharm. Bull. (Tokyo) 41: 214-216 (1993) https://doi.org/10.1248/cpb.41.214
  11. Chen C, Chou CK, Lee SD, Wang JC, Yeh SF. Active compounds from Saussurea lappa Clarks that suppress hepatitis B virus surface antigen gene expression in human hepatoma cells. Antivir. Res. 27: 99-109 (1995) https://doi.org/10.1016/0166-3542(94)00083-K
  12. Wedge DE, Galindo JC, Marcias FA. Fungidal activity of natural and synthetic sesquiterpene lactone analogs. Phytochemistry 53: 747-757 (2000) https://doi.org/10.1016/S0031-9422(00)00008-X
  13. Cho JY, Baik KU, Jung JH, Park MH. In vitro anti-inflammatory effects of cynaropicrin, a sesquiterpene lactone, from Saussurea lappa. Eur. J. Pharmacol. 398: 399-407 (2000) https://doi.org/10.1016/S0014-2999(00)00337-X
  14. Matsuda H, Toguchida I, Ninomiya K, Kageura T, Morikawa T, Yoshikawa M. Effects of sesquiterpenes and amino acid-sesquiterpene conjugates from the roots of Saussurea lappa on inducible nitric oxide synthase and heat shock protein in lipopolysaccharide- activated macrophages. Bioorg. Med. Chem. Lett. 11: 709-715 (2003) https://doi.org/10.1016/S0968-0896(02)00471-6
  15. Jeong SJ, Itokawa T, Shibuya M, Kuwano M, Ono M, Higuchi R, Miyamoto T. Costunoide, a sesquiterpene lactone from Saussurea lappa, inhibits the VEGFR KDR/Flk-1 signaling pathway. Cancer Lett. 187: 129-133 (2002) https://doi.org/10.1016/S0304-3835(02)00361-0
  16. Ko SG, Koh SH, Jun CY, Nam CG, Bae HS, Shin MK. Induction of apoptosis Saussurea lappa by and Pharbits nil on AGS gastric cancer cells. Biol. Pharm. Bull. 27: 1604-1610 (2004) https://doi.org/10.1248/bpb.27.1604
  17. Ko SG, Kim HP, Jin DH, Bae HS, Kim SH, Park CH, Lee JW. Saussurea lappa induces G2-growth arrest and apoptosis in AGS gastric cancer cells. Cancer Lett. 220: 11-19 (2005) https://doi.org/10.1016/j.canlet.2004.06.026
  18. Bocca C, Gabriel L, Bozzo F, Miglietta A. A sesquiterpene lactone, costunolide, interacts with microtubule protein and inhibits the growth of MCF-7 cells. Chem. Biol. Interact. 147: 79-86 (2004) https://doi.org/10.1016/j.cbi.2003.10.008
  19. Choi SH, Im E, Kang HK, Lee JH, Kwak HS, Bae YT, Park HJ, Kim ND. Inhibitory effects of costunolide on the telomerase activity in human breast carcinoma cells. Cancer Lett. 227: 153-162 (2005) https://doi.org/10.1016/j.canlet.2005.01.011
  20. Choi JH, Seo BR, Seo SH, Lee KT, Park JH, Park HJ, Choi JW, Itoh Y, Miyamoto K. Costunolide induces differentiation of human leukemia HL-60 cells. Arch. Pharm. Res. 25: 480-484 (2002) https://doi.org/10.1007/BF02976606
  21. Hengartner MO. The biochemistry of apoptosis. Nature 407: 770-777 (2000) https://doi.org/10.1038/35037710
  22. Korsmeyer SJ. Regulators of cell death. Trends Genet. 11: 101-105 (1995) https://doi.org/10.1016/S0168-9525(00)89010-1
  23. Debatin KM. Apoptosis pathways in cancer and cancer therapy. Cancer Immunol. Immun. 53: 153-159 (2004) https://doi.org/10.1007/s00262-003-0474-8
  24. Ramachandran A, Madesh M, Balasubramanian KA. Apoptosis in the intestinal epithelium: Its relevance in normal and pathophysiology conditions. J. Gastoenterol. Hepatol. 15: 109-120 (2000) https://doi.org/10.1046/j.1440-1746.2000.02059.x
  25. Kim DG. Apoptosis and gastrointestinal disease. Korean J. Gastroenterol. 39: 79-87 (2002)
  26. Haldar S, Chintapalli J, Croce CM. Taxol induces bcl-2 phosphorylation and death of prostate cancer cells. Cancer Res. 56: 1253-1255 (1996)
  27. Lowe SW, Lin AW. Apoptosis in cancer. Carcinogenesis 21: 485-495 (2000) https://doi.org/10.1093/carcin/21.3.485
  28. Schmitt CA, Lowe SW. Apoptosis and therapy. J. Pathol. 187: 127-137 (1999) https://doi.org/10.1002/(SICI)1096-9896(199901)187:1<127::AID-PATH251>3.0.CO;2-T
  29. Denizot F, Lang R. Rapid colorimetric assay for cell growth and survival modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J. Immunol. Methods 89: 271-277 (1986) https://doi.org/10.1016/0022-1759(86)90368-6
  30. Kim EJ, Shin HK, Cho JS, Lee SK, Won MH, Kim HW, Park JHY. Trans-10,cis-12 conjugated linoleic acid inhibits the G1-S cell cycle progression in DU145 human prostate carcinoma cells. J. Med. Food 9: 293-299 (2006) https://doi.org/10.1089/jmf.2006.9.293
  31. Jung JI, Lim SS, Choi HJ, Cho HJ, Shin HK, Kim EJ, Chung WY, Park KK, Park JHY. Isoliquiritigenin induces apoptosis by depolarizing mitochondrial membranes in prostate cancer cells. J. Nutr. Biochem. 17: 689-696 (2006) https://doi.org/10.1016/j.jnutbio.2005.11.006
  32. Kim EJ, Houlthuizen PE, Park HS, Ha YL, Jung KC, Park JHY. Trans-10,cis-12 conjugated linoleic acid inhibits Caco-2 colon cancer cell growth. Am. J. Physiol. Gastrointest. Liver Physiol. 283: G357-G367 (2002) https://doi.org/10.1152/ajpgi.00495.2001
  33. Kim EJ, Park SY, Hong J, Shin M, Lim SS, Shin HK, Park JHY. Inhibitory effect of the methanolic extract of Symphyocladia latiuscula on the growth of HT-29 human colon cancer cells. J. Korean Soc. Food Sci. Nutr. 36: 431-438 (2007) https://doi.org/10.3746/jkfn.2007.36.4.431
  34. Yu Z, Li W. Induction of apoptosis by puerarin in colon cancer HT-29 cells. Cancer Lett. 238: 53-60 (2006) https://doi.org/10.1016/j.canlet.2005.06.022
  35. Cory S, Huang DC, Adams JM. The Bcl-2 family: Roles in cell survival and oncogenesis. Oncogene 22: 8590-8607 (2003) https://doi.org/10.1038/sj.onc.1207102
  36. Bouillet P, Strasser A. BH3-only proteins-evolutionarily conserved proapoptotic Bcl-2 family members essential for initiating programmed cell death. J. Cell Sci. 115: 1567-1574 (2002)
  37. Oltvai ZN, Milliman CL, Korsmeyer SJ. Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell 74: 609-619 (1993) https://doi.org/10.1016/0092-8674(93)90509-O
  38. Kim EJ, Lee YJ, Shin HK, Park JHY. A study on the mechanisms by which the aqueous extract of Inonotus obliquus induces apoptosis and inhibits proliferation in HT-29 human colon cancer cells. J. Korean Soc. Food. Sci. Nutr. 35: 516-523 (2006) https://doi.org/10.3746/jkfn.2006.35.5.516
  39. Kim EJ, Lee YJ, Shin HK, Park JHY. Induction of apoptosis by the aqueous extract of Rubus coreanum in HT-29 human colon cancer cells. Nutrition 21: 1141-1148 (2005) https://doi.org/10.1016/j.nut.2005.02.012
  40. Oliver FJ, de la Rubia G, Rolli V, Ruiz-Ruiz MC, de Murcia G, Murcia JM. Importance of poly(ADP-ribose) polymerase and its cleavage in apoptosis. Lesson from an uncleavable mutant. J. Biol. Chem. 273: 33533-33539 (1998) https://doi.org/10.1074/jbc.273.50.33533