동아시아식생활학회지 (Journal of the East Asian Society of Dietary Life)

  • 제23권6호
  • /
  • Pages.723-732
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  • 2013
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  • 1225-6781(pISSN)
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  • 2288-8802(eISSN)
동아시아식생활학회 (The East Asian Society of Dietary Life)
권호 표지

복분자의 유방암 세포 사멸 및 사멸 유전자 발현 유도 효과

Inducing Effects of Rubus coreanus on Cell Death and Apoptotic Gene Expressions in Human Breast Cancer Cells

  • 김희정 (덕성여자대학교 자연과학대학 식품영양학과) ;
  • 강금지 (덕성여자대학교 자연과학대학 식품영양학과)
  • Kim, Hee-Jung (Dept. of Food & Nutrition, College of Natural Science, Duksung Women's University) ;
  • Kang, Keum-Jee (Dept. of Food & Nutrition, College of Natural Science, Duksung Women's University)
  • 투고 : 2013.09.17
  • 심사 : 2013.12.30
  • 발행 : 2013.12.31
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초록

We investigated the inducing effects of Rubus coreanus extract (RCE) on apoptosis and its related gene expressions in human breast cancer cells. MDA-MB-231 cells were cultured in the presence of 0, 200, 300, and $400{\mu}g/mL$ RCE for 24h. MTT assay demonstrated that relative cell viability measured a decrease in a dose-dependent manner (p<0.05). This dependency was also found in the increasing levels of cell death by a dual staining with Hoechst 33322 and propidium iodide (p<0.05). These close associations was also observed by different stages of apoptotic processes, as shown by an Apoptosis Detection Kit. To determine whether the alterations in such cell activities obtained above cause the induction of apoptotic genes, PT-PCR was performed expressions of both Bcl-2 and Bax mRNAs. The Bcl-2/Bax ratio which is an important indicator of apoptosis, was found to have significantly decreased dose dependence (p<0.05). Western blot analysis also demonstrated that Caspase-3 significantly increases in a dose-dependent manner (p<0.05) in addition to similar alterations of other proteins examined. Taken these results together, the ethanolic RCE used induces a reduction in cell viability along with increased membrane permeability. This leads to a precautious apoptotic process and, subsequently, cell death through the apoptotic pathway involving Bax and Caspase-3 in human breast cancer MDA-MB-231 cells.

키워드

참고문헌

  1. Aiyer HS, Warri AM, Woode DR, Hilakivi CL, Clarker R (2012) Influence of berry polyphenols on receptor signaling and cell death pathways: Implications for breast cancer prevention. J Agric Food Chem 60: 5693-5708. https://doi.org/10.1021/jf204084f
  2. American Cancer Society (2013) Breast cancer: Facts & figures 2013-2014. American Cancer Society Inc., Atlanta. pp 4-11.
  3. Beak BK, Lim CW, Lee EY, Whang IS, Kwon HN, Lee HK, Lee SI, Lee HJ, Jeon BH (2004) Effects of raspberry wine on testosterone level of Sprague Dawley rats. Korean J Oriental Physiol Pathol 18: 1007-1013.
  4. Belloc F, Dumain P, Boisseau MR, Jalloustre C (1994) A flow cytometric method using Hoechst 33342 and propidium iodide for simultaneous cell cycle analysis and apoptosis determination in unfixed cells. Cytometry 17: 59-65. https://doi.org/10.1002/cyto.990170108
  5. Bhandary B, Lee GH, Marahatta A, Lee HY, Kim SY, So BO, Kwon JW, Song JY, Lee HK, Kim HR, Chae SW, Chae HJ (2012) Water extracts of immature Rubus coreanus regulate lipid metabolism in liver cells. Biol Pharm Bull 35: 1907- 1913. https://doi.org/10.1248/bpb.b12-00022
  6. Boivin D, Blanchette M, Barrette S, Moghrabi A, Beliveau R (2007) Inhibition of cancer cell proliferation and suppression of $TNF\alpha$-induced activation of NF-${\kappa}B$ by edible berry juice. Anticancer Res 27: 937-948.
  7. Choi C, Lee H, Lim H, Park S, Lee J, Do S (2012) Effect of Rubus coreanus extracts on diabetic osteoporosis by simultaneous regulation of osteoblasts and asteoclasts. Menopause 19: 1043-51. https://doi.org/10.1097/gme.0b013e31824b1cc5
  8. Chen WF, Huang MH, Tzang CH, Yang M, Song MS (2003) Inhibitory actions of genistein in human breast cancer (MCF-7) cells. Biochim Biophys Acta 1638: 187-196. https://doi.org/10.1016/S0925-4439(03)00082-6
  9. Coates EM, Popa G, Gill CI, Mccann MJ, Dougall GJ, Stewart D, Rowland I (2007) "Colon-available" raspberry polyphenols exhibit anti-cancer effects on in vitro models of colon cancer. J Carcinog 6: 4-16. https://doi.org/10.1186/1477-3163-6-4
  10. Del Poeta G, Venditti A, Del Principe MI (2003) Amount of spontaneous apoptosis detected by Bax/Bcl-2 ratio predicts outcome in acute myeloid leukemia. Blood 101: 2125-2131. https://doi.org/10.1182/blood-2002-06-1714
  11. Gross A, McDonnell JM, Korsmeyer SJ (1999) Bcl-2 family members and the mitochondria in apoptosis. Genes 13: 1899-1911. https://doi.org/10.1101/gad.13.15.1899
  12. Hidayat T, Yuliarto S, Barlianto W, Sujuti H, Chandra Kusuma HMS (2013) Increase of Bcl-2/Bax ratio corelated with decrease of lymphocyte apoptosis: A study in the bronchiolus and lung of asthmatic mice. Int J Med Med Sci 5: 191-197.
  13. Jakobek L, Seruga M, Seruga B, Novak L (2011) Phenolic compound composition and antioxidant activity of fruits of Rubus and Prumus species from Croatia. Int J Food Sci Technol 44: 860-868.
  14. Jeong JH, Jung H, Lee SR, Lee HJ, Hwang KT, Kim TY (2010) Anti-oxidant, anti-proliferative and anti-inflammatory activities of the extracts from black raspberry fruits and wine. Food Chemistry 123: 338-344. https://doi.org/10.1016/j.foodchem.2010.04.040
  15. Ju HK, Cho EJ, Jang MH, Lee YY, Hong SS, Park JH, Kwon SW (2009) Characterization of increased phenolic compounds from fermented bokbunja and related antioxidant activity. J Pharm Biomed Anal 49: 820-827. https://doi.org/10.1016/j.jpba.2008.12.024
  16. Jung KA, Han D, Kwon EK, Lee CH, Kim YE (2007) Antifatigue effect of Rubus coreanus Miquel extract in mice. J Med Food 10: 689-693. https://doi.org/10.1089/jmf.2006.006
  17. Kastan MB, Bartek J (2004) Cell-cycle checkpoints and cancer. Nature 432: 316-323. https://doi.org/10.1038/nature03097
  18. Kim EJ, Lee YJ, Shin HK, Park JH (2005) Induction of apoptosis by the aqueous extract of Rubus coreanus in HT-29 human colon cancer cells. Nutr 21: 1141-1148. https://doi.org/10.1016/j.nut.2005.02.012
  19. Kim HH, Choi PH, Yoo JS, Jeon H, Chae BS, Park JS, Kim SH, Shin TY (2012a) Ripe fruit of Rubus coreanus inhibits mast cell mediated allergic inflammation. Int J Mol Med 29: 303-310.
  20. Kim YR, Kim JN, Lee SM, Lee HA, Park SH, Kim YS, Kim JH (2012b) Chemopreventive effects of Rubus coreanus Miquel on prostate cancer. Biosci Biotech Biochem 76: 737-744. https://doi.org/10.1271/bbb.110857
  21. Lee SN, Kang KJ (2010) The effect of blueberry extract in gene expressions related to apoptosis in human breast cancer MCF-7 cells. J East Asian Soc Dietary Life 20: 30-36.
  22. Liu D, Chen Z (2013) The effect of curcumin on breast cancer cells. J breast Cancer 16: 133-137. https://doi.org/10.4048/jbc.2013.16.2.133
  23. Liu Y, Peterson DA, Kimura H, Schubert D (1997) Mechanism of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction. J Neurochem 69: 581-593.
  24. Mackey TJ, Borkowski A, Amin P, Jacobs SC, Kyprianou N (1998) bcl-2/bax ratio as a predictive marker for therapeutic response to radiotherapy in patients with prostate cancer. Urology 52: 1085-1090. https://doi.org/10.1016/S0090-4295(98)00360-4
  25. Ministry of Health and Welfare (2012) and National Cancer Center. Cancer Facts & Figures 2012 in the Republic of Korea, Gyeonggi-Do. pp 8-15.
  26. 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
  27. Nam MK, Kang KJ (2013) The effect of red cabbage extract on the apoptosis in human breast cancer MDA-MB-231 cells. J Korean Soc Food Sci Nutr 42: 8-16. https://doi.org/10.3746/jkfn.2013.42.1.008
  28. Navindra P, Seeram NP (2008) Berry fruits for cancer prevention: Current status and future prospects. J Agric Food Chem 56: 630-635. https://doi.org/10.1021/jf072504n
  29. Park YK, Choi SH, Kim SH, Jang YS, Han JG, Chung HG (2008) Functional composition and antioxidant activity from the fruit of Rubus coreanus according to cultivars. J Korean Wood Sci Technol 36: 102-109.
  30. Pierce JP, Natarajan L, Caan BJ, Parker BA, Greenberg ER, Flatt SW, Rock CL, Kealey S, Al-Delaimy WK, Bardwell WA, Carlson RW, Emond JA, Faerber S, Gold EB, Hajek RA, Hollenbach K, Jones LA, Karanja N, Madlensky L, Marshall J, Newman VA, Ritenbaugh C, Thomson CA, Wasserman L, Stefanick ML (2007) Influence of a diet very high in vegetables, fruit and fiber and low in fat on prognosis following treatment for breast cancer: the women's healthy eating and living randomized trial. JAMA 298: 289-298. https://doi.org/10.1001/jama.298.3.289
  31. Ravoor S, Vadhaam MV, Agil F, Gupta RC (2012) Inhibition of estrogen-mediated mammary tumorigenesis by blueberry and black raspberry. J Agric Food Chem 60: 5547-5555. https://doi.org/10.1021/jf205325p
  32. Reed JC (1994) Bcl-2 and the regulation of programmed cell death. J Cell Biol 124: 1-6. https://doi.org/10.1083/jcb.124.1.1
  33. Reynolds CP, Black AT, Woody JN (1986) Sensitive method for detecting viable cells seeded into bone marrow. Cancer Res 46: 5878-5881.
  34. Seeram NP, Herber D (2007) Impact of berry phytochemicals on human health: Effects beyond antioxidation. ACS Symposium Series 956: 326-336.
  35. Suganya DP, Saravana KM, Mohan SD (2011) Evaluation of antiproliferative activity of red sorghum bran anthocyanin on a human breast cancer cell line (MCF-7). Int J Breast Cancer 2011: 1-6.
  36. Vogelstein BD, Lane AJ (2000) Surfing the p53 network. Nature 408: 307-310. https://doi.org/10.1038/35042675
  37. Yang E, Zha J, Jockel J, Boise LH, Thompson CB, Kosmeyer SJ (1995) Bad, a heterodimeric partner for bcl-XL and bcl-2, displaces bax and promotes cell death. Cell 80: 285- 291. https://doi.org/10.1016/0092-8674(95)90411-5
  38. Yang J, Liu X, Bhalla K, Kim CN, Ibrado AM, Cai J (1997) Prevention of apoptosis by bcl-2: release of cytochrome c from mitochondria blocked. Science 275: 1129-1132. https://doi.org/10.1126/science.275.5303.1129
  39. Yang HM, Oh SM, Lim SS, Shin HK, Oh YS, Kim JK (2008) Antiinflammatory activities of Rubus coreanus depend on the degree of fruit ripening. Phytother Res 22: 102-107. https://doi.org/10.1002/ptr.2274
  40. Yang XH, Edgerton S, Thor AD (2005) Reconstitution of caspase- 3 sensitizes MCF-7 breast cancer cells to radiation therapy. Int J Oncol 26: 1675-1680.
  41. Yoon I, Cho JY, Kuk JH, Wee JH, Jang MY, Ahn TH (2001) Identification and activity of antioxidative compounds from Rubus coreanum fruit. Korean J Food Sci Technol 34: 898- 904.
  42. Zhou J, Zhong Y (2004) Breast cancer immunotherapy. Cell Mol Immunol 1: 247-255.