Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
권호 표지
DOI QR코드

DOI QR Code

Inhibitory Effect of the Hexane Extract of Saussurea lappa on the Growth of LNCaP Human Prostate Cancer Cells

목향 헥산추출물의 LNCaP 전립선암세포 증식 억제 효과

  • Park, So-Young (Dept. of Food Sciences and Nutrition, Hallym University) ;
  • Kim, Eun-Ji (Center for Efficacy Assessment and Development of Functional Foods and Drugs, Hallym University) ;
  • Lim, Do-Young (Dept. of Food Sciences and Nutrition, Hallym University) ;
  • Kim, Jong-Sang (Dept. of Animal Science and Biotechnology, Kyungpook National University) ;
  • Lim, Soon-Sung (Center for Efficacy Assessment and Development of Functional Foods and Drugs, Hallym University) ;
  • Shin, Hyun-Kyung (Dept. of Food Sciences and Nutrition, Hallym University) ;
  • Yoon Park, Jung-Han (Dept. of Food Sciences and Nutrition, Hallym University)
  • 박소영 (한림대학교 식품영양학과) ;
  • 김은지 (한림대학교 식의약품의 효능평가 및 기능성소재개발센터) ;
  • 임도영 (한림대학교 식품영양학과) ;
  • 김정상 (경북대학교 동물공학과) ;
  • 임순성 (한림대학교 식의약품의 효능평가 및 기능성소재개발센터) ;
  • 신현경 (한림대학교 식품영양학과) ;
  • 윤정한 (한림대학교 식품영양학과)
  • Published : 2008.01.31
Download PDF
⟨ Previous Next ⟩

Abstract

Saussurea lappa (SL) has been used to reduce abdominal pain and tenesmus in traditional oriental medicine. SL and major compounds of SL, sesquiterpene lactones, have been suggested to possess various biological effects, including anti-tumor, anti-ulcer, anti-inflammatory, anti-viral and cardiotonic activities. Recently, it has been reported that ethanol extracts from roots of SL have antiproliferative effects on gastric cancer cells. To explore the possibility that SL has chemopreventive effects on prostate cancer, we examined whether the hexane extract of SL (HESL) inhibits the growth of LNCaP human prostate cancer cells. Cells were incubated with various concentrations ($0{\sim}4$ mg/L) of HESL in DMEM/F12 containing 5% charcoal stripped fetal bovine serum. HESL substantially decreased viable cell numbers and induced apoptosis of LNCaP cells in dose-dependent manners. HESL increased the levels of cleaved caspase-8, -9, -7 and -3, and poly (ADP-ribose) polymerase. HESL increased the levels of the pro-apoptotic Bak and truncated-Bid proteins whereas it had no effect on the anti-apoptotic Bcl-2, Bcl-xL, or Mcl-1. The present results indicate that HESL inhibits the growth of human prostate cancer cells by inducing apoptosis, which involves the activation of the caspase cascades.

목향은 국화과에 속한 다년생 식물인 Saussurea lappa의 뿌리로서 한의학에서는 구토, 설사 및 염증치료 등에 사용되고 있다. 목향 추출물 및 그 성분들은 항균 작용, 항염증 작용, 혈관생성 억제 효능 등을 지니고 있으며 위암과 대장암의 세포증식을 억제하는 것으로 알려져 있다. 그러나 현재까지 전립선암에 대한 목향의 효과는 연구된 바 없다. 본 연구에서는 목향이 전립선 암세포에 미치는 영향을 조사하기 위해 인간의 전립선에서 유래한 암세포인 LNCaP 세포의 증식과 apoptosis에 미치는 영향을 조사하였다. 목향 헥산추출물을 LNCaP 세포 배양액에 여러 농도($0{\sim}4$ mg/L)로 첨가하여 세포의 증식에 미치는 영향을 조사하였다. 목향 헥산추출물의 농도가 증가할수록 LNCaP 세포의 증식은 현저하게 감소하였고 apoptosis는 증가함을 관찰하였다. 목향 헥산추출물이 LNCaP 세포의 apoptosis 일으키는 기전을 연구하기 위하여 목향 헥산추출물을 첨가하고 세포를 48시간 배양한 후 cell lysate를 얻어 Western blot을 실시하였다. Apoptosis 과정에 작용하는 중요한 단백질 중 하나인 Bcl-2 family 단백질 중 pro-apoptotic Bcl-2 단백질인 Bak와 BH3 only Bcl-2 단백질인 truncated-Bid의 단백질 수준은 목향 헥산 추출물에 의해 유의적으로 증가한 반면 anti-apoptotic Bcl-2 단백질인 Bcl-2, Bcl-xL 및 Mcl-1 단백질 수준은 변하지 않았다. 또한 apoptosis를 집행하는 caspase의 활성 형태인 cleaved caspase-8, -9, -7, -3의 단백질 수준이 목향 헥산추출물의 처리에 의해 증가하였고 caspase-3의 표적 단백질 중 하나인 PARP의 불활성 형태인 cleaved PARP의 단백질 수준도 현저하게 증가하였다. 이 결과들은 목향 헥산 추출물이 LNCaP 세포의 apoptosis를 유도함으로써 전립선 암세포의 증식을 억제함을 보여주는 것이며 목향 헥산추출물에 의한 apoptosis 유도는 caspase 활성 증가와 Bak 및 t-Bid 단백질의 증가에 의한 것임을 제시한다. 따라서 앞으로 항암효과를 나타내는 성분의 동정 및 동물실험을 통하여 좀 더 면밀한 기전 연구가 수행된다면 목향 헥산추출물은 화학적 암예방 물질이나 치료제로 개발될 수 있을 것으로 사료된다.

Keywords

References

  1. Jeon YJ, Lee HS, Yeon SY, Ko JH, An KM, Yu SW, Kang JH, Hwang BY, Kin TY. 2005. Inhibitory effect of dehydrocostuslactone isolated from Sussureae Radix on CDK2 activity. Kor J Pharmacogn 36: 97-101
  2. Jung NH, Ha JY, Min KR, Shibata F, Nakagawa H, Kang SS, Chang IM, Kim YS. 1998. Reynosis from Saussurea lappa as inhibitor on CINC-1 induction in LPS-stimulated NRK-52E cells. Planta Med 64: 454-456 https://doi.org/10.1055/s-2006-957480
  3. Kang SH, Kim JS, Chi HJ, Chang SY, Ha KW. 1999. Isolation and quantitative determination of costunolide from saussurea root. Kor J Pharmacogn 30: 48-53
  4. Wedge DE, Galindo JC, Macias FA. 2000. Fungcidal activity of natural and synthetic sesquiterpene lactone analogs. Phytochem 53: 747-757 https://doi.org/10.1016/S0031-9422(00)00008-X
  5. Park HJ, Jung WT, Basnet P, Kadota S, Namba T. 1996. Syringin 4-O-$\beta$ glucoside, a new phenylpropanoid glycoside, and costunolide, a nitric oxide synthase inhibitor, from the stem bark of Magnolia sieboldii. J Nat Prod 59: 1128-1130 https://doi.org/10.1021/np960452i
  6. Jeong SJ, Ltokawa T, Shibuya M, Kuwano M, Ono H, Huguchi R, Miyamoto T. 2002. Costunolide, a sesquiterpene lactone from Saussurea lappa, inhibits the VEGFR KDR ⁄Flk-1 signaling pathway. Cancer Lett 187: 129-133 https://doi.org/10.1016/S0304-3835(02)00361-0
  7. Ko SG, Koh SH, Jun CY, Nam CG, Bae HS, Shin MK. 2004. Induction of apoptosis by Saussurea lappa and Pharbitis nil on AGS gastric cancer cells. Biol Pharm Bull 27: 1604-1610 https://doi.org/10.1248/bpb.27.1604
  8. Ko SG, Kim HP, Jim DH, Bae HS, Kim SH, Park CH, Lee JW. 2005. Saussurea lappa induces G2-growth arrest and apoptosis in AGS gastric cancer cells. Cancer Lett 220: 11-19 https://doi.org/10.1016/j.canlet.2004.06.026
  9. Lee MG, Lee KT, Chi SG, Park JH. 2001. Costunolide induces apoptosis by ROS-mediated mitochondrial permeability transition and cytochrome c release. Biol Pharm Bull 24: 303-306 https://doi.org/10.1248/bpb.24.303
  10. Mori H, Kawamori T, Tanaka T, Ohnish M, Yamahara J. 1994. Chemopreventive effect of costunolide, a constituent of oriental medicine, on azoxymethane-induced intestinal carcinogenesis in rats. Cancer Lett 83: 171-175 https://doi.org/10.1016/0304-3835(94)90315-8
  11. Chen CN, Huang HH, Wu CL, Lin PC, Hsu JT, Hsieh HP, Chuang SE, Lai GM. 2007. Isocostunolide, a sesquiterpene lactone, induces mitochondrial membrane depolarization and caspase-dependent apoptosis in human melanoma cells. Cancer Lett 246: 237-252 https://doi.org/10.1016/j.canlet.2006.03.004
  12. Martin KR. 2007. Using nutrigenomics to evaluate apoptosis as a preemptive target in cancer prevention. Curr Cancer Drug Targets 7: 438-446 https://doi.org/10.2174/156800907781386650
  13. Hengartner MO. 2000. The biochemistry of apoptosis. Nature 407: 770-777 https://doi.org/10.1038/35037710
  14. Debatin KM. 2004. Apoptosis pathways in cancer and cancer therapy. Cancer Immunol Immunother 53: 153-159 https://doi.org/10.1007/s00262-003-0474-8
  15. Im SS, Kim JR, Lim HA, Jang CH, Kim YK, Konishi T, Kim EJ, Park JHY, Kim JS. 2007. Induction of detoxifying enzyme by seszuiterpenes present in Inula helenium. J Med Food 10: 503-510 https://doi.org/10.1089/jmf.2006.209
  16. Denizot F, Lang R. 1986. 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 https://doi.org/10.1016/0022-1759(86)90368-6
  17. Hodge GL, Flower R, Han P. 1999. Optimal storage conditions for preserving granulocyte viability as monitored by Annexin V binding in whole blood. J Immunol Methods 225: 27-38 https://doi.org/10.1016/S0022-1759(99)00025-3
  18. Herault O, Colombat P, Domenech J, Degenne M, Bremone JL, Sensebe L, Bernard MC, Binet C. 1999. A rapid single-laser flow cytometric method for discrimination of early apoptotic cells in a heterogeneous cell population. Br J Haematol 104: 530-537 https://doi.org/10.1046/j.1365-2141.1999.01203.x
  19. Cowell JK, Franks LM. 1980. A rapid method for accurate DNA measurements in single cells in situ using a simple microfluorimeter and Hoechst 33258 as a quantitative fluorochrome. J Histochem Cytochem 28: 206-210 https://doi.org/10.1177/28.3.6153398
  20. Jeon YJ, Lee HS, Yeon SY, Ko JH, An KM, Yu SW, Kang JH, Hwang BY, Kin TY. 2005. Inhibitory effect of dehydrocostuslactone isolated from Sussureae Radix on CDK2 activity. Kor J Pharmacogn 36: 97-101
  21. Bocca C, Gabriel L, Bozzo F, Miglietta A. 2004. A sisquiterpene lactone, costunolide, interacts with microtubule protein and inhibits the growth of MCF-7 cells. Chem Biol Interact 147: 79-86 https://doi.org/10.1016/j.cbi.2003.10.008
  22. Park HJ, Kwon SH, Han YN, Choi JW, Miyamoto K, Lee SH, Lee KT. 2001. Apoptosis- inducing costunolide and a novel acyclic monoterpene from the stem bark of Magnolia sieboldii. Arch Pharm Res 24: 342-348 https://doi.org/10.1007/BF02975104
  23. Moon BK. 2002. Anesthesiology and apoptosis. J Kor Anesthesiology 42: 563-574 https://doi.org/10.4097/kjae.2002.42.5.563
  24. Banasiak KJ, Sia Y, Haddad GG. 2000. Mechanisms underlying hypoxia-induced neuronal apoptosis. Prog Neurobiol 62: 215-249 https://doi.org/10.1016/S0301-0082(00)00011-3
  25. Amundson SA, Myers TG, Fornace AJ Jr. 1998. Roles for p53 in growth arrest and apoptosis: putting on the brakes after genotoxic stress. Oncogene 17: 3287-3299 https://doi.org/10.1038/sj.onc.1202576
  26. Rokhlin OW, Bishop GA, Hostager BS, Waldschmidt TJ, Sidorenko SP, Pavloff N, Kierfer MC, Umansky SR, Cohen MB. 1997. Fas-mediated apoptosis in human prostatic carcinoma cell lines. Cancer Res 57: 1758-1768
  27. Kim EJ, Park SY, Hong J, Shin M, Lim SS, Shin HK, Park JHY. 2007. 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 https://doi.org/10.3746/jkfn.2007.36.4.431
  28. Vermeulen K, Van Bockstaele DR, Berneman ZN. 2005. Apoptosis: mechanisms and relevance in cancer. Ann Hematol 84: 627-639 https://doi.org/10.1007/s00277-005-1065-x
  29. Lemasters JJ, Quian T, Bradham CA, Brenner DA, Cascio WE, Trost LC, Nishimura Y, Nieminen AL, Herman B. 1999. Mitochondrial dysfunction in the pathogenesis of necrotic and apoptotic cell death. J Bioenerg Biomembr 31: 305-319 https://doi.org/10.1023/A:1005419617371
  30. Yamaguchi H, Chen J, Bhalla K, Wang HG. 2004. Regulation of Bax activation and apoptotic response to microtubule-damaging agents by p53 transcription-dependent and-independent pathways. J Biol Chem 279: 39431-39437 https://doi.org/10.1074/jbc.M401530200
  31. Licholson DW, Thornberry NA. 1997. Caspases: Killer pergeases. Trend Biochem Sci 22: 299-306 https://doi.org/10.1016/S0968-0004(97)01085-2
  32. Ho PK, Hawkins CJ. 2005. Mammalin initiator apoptotic caspases. FEBS J 272: 5436-5453 https://doi.org/10.1111/j.1742-4658.2005.04966.x
  33. Budihardjo I, Oliver H, Lutter M, Luo X, Wang X. 1999. Biochemical pathways of caspase activation during apoptosis. Annu Rev Cell Dev Biol 15: 269-290 https://doi.org/10.1146/annurev.cellbio.15.1.269
  34. Kim EJ, Park SY, Shin HK, Kwon DY, Surh YJ, Park JHY. 2007. Activation of caspase-8 contributes to 3,3'-diindolylmethane-induced apoptosis in colon cancer cells. J Nutr 137: 31-36
  35. Oliver FJ, de la Rubia G, Folli V, Ruiz-Ruiz MC, de Murcia G, Murcia JM. 1998. Importance of poly(ADP-ribose) polymerase and its cleavage in apoptosis. Lesson form an uncleavable mutant. J Biol Chem 273: 33533-33539 https://doi.org/10.1074/jbc.273.50.33533
  36. Pulmmer ER. 2006. Inhibition of poly(ADP-ribose) polymerase in cancer. Curr Opin Pharmacol 6: 364-368 https://doi.org/10.1016/j.coph.2006.02.004
  37. Ratnam K, Low JA. 2007. Current development of clinical inhibitors of poly(ADP-ribose) polymerase in oncology. Clin Cancer Res 13: 1383-1388 https://doi.org/10.1158/1078-0432.CCR-06-2260

Cited by

  1. induction of Apoptosis in Human Cancer Cells with Compositae Extracts vol.41, pp.5, 2012, https://doi.org/10.3746/jkfn.2012.41.5.584
  2. The Effect of Hwangryunhaedoktang on Proliferations of Various Human Cancer Cells vol.27, pp.1, 2013, https://doi.org/10.7778/jpkm.2013.27.1.059
  3. Resveratrol Induces Apoptosis in Primary Human Prostate Cancer Cells vol.39, pp.8, 2010, https://doi.org/10.3746/jkfn.2010.39.8.1119
  4. Antioxidant and antiproliferating effects of Setaria italica, Panicum miliaceum and Sorghum bicolor extracts on prostate cancer cell lines vol.23, pp.7, 2016, https://doi.org/10.11002/kjfp.2016.23.7.1033
  5. 레스베라트롤에 의한 인간 암세포주, A549와 SKOV3의 p53의존적 Apoptosis 유발 vol.44, pp.2, 2016, https://doi.org/10.4014/mbl.1512.12013
  6. Effect of Aucklandia lappa Decne Extract on Antioxidant vol.37, pp.6, 2020, https://doi.org/10.12925/jkocs.2020.37.6.1545