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Antioxidant Activities of Perilla frutescens Britton Seed Extract and Its Inhibitory Effects against Major Characteristics of Cancer Cells

들깨 추출물의 항산화 활성과 암세포 기본 특성에 대한 억제 효과

  • Kim, Sinae (Department of Food and Nutrition, Chungbuk National University) ;
  • Song, Boram (Department of Food and Nutrition, Chungbuk National University) ;
  • Ju, Jihyeung (Department of Food and Nutrition, Chungbuk National University)
  • 김시내 (충북대학교 식품영양학과) ;
  • 송보람 (충북대학교 식품영양학과) ;
  • 주지형 (충북대학교 식품영양학과)
  • Received : 2014.11.04
  • Accepted : 2014.12.23
  • Published : 2015.02.28

Abstract

The aim of the study was to investigate the antioxidant activities of ethanol extract of perilla seed (PSE) and its inhibitory effects against major characteristics of cancer cells, such as unrestricted growth and activated metastasis in vitro. The total polyphenol and flavonoid levels of PSE were 222.6 mg gallic acid equivalent/100 g and 285.7 mg quercetin equivalent/100 g, respectively. The radical scavenging activity and ferric reducing antioxidant power of PSE at concentration of 87.5 to $350{\mu}g/mL$ were 24~45% and 28~62%, respectively. Treatment of HCT116 colorectal carcinoma cells and H1299 non-small cell lung carcinoma cells with PSE dose-dependently inhibited growth by 18~94% (at concentration range of 87.5 to $350{\mu}g/mL$) and completely abolished colony formation (at concentration of $175{\mu}g/mL$). PSE was also effective in inhibiting migration of H1299 cells (by 30~37% at concentration range of 87.5 to $350{\mu}g/mL$) and adhesion of both HCT116 and H1299 cells (by 14~16% at concentration of $350{\mu}g/mL$). These results indicate that PSE exerts antioxidant and anti-cancer activities in vitro. It needs to be determined whether or not similar effects can be reproduced in vivo.

본 연구에서는 들깨 에탄올 추출물의 항산화 성분 함량 및 활성을 측정하고 들깨 추출물이 암세포의 주요 특징인 성장, colony 형성, 이동, 부착성 등에 미치는 영향을 in vitro 수준에서 분석하고자 하였다. 들깨의 총 폴리페놀 함량과 총 플라보노이드 함량은 각각 222.6 mg gallic acid equivalent/100 g과 285.7 mg quercetin equivalent/100 g으로 측정되었다. 들깨 추출물($87.5{\sim}350{\mu}g/mL$)의 DPPH radical 소거 활성은 24~45%, 철 환원력은 28~62%였다. 한편 $87.5{\sim}350{\mu}g/mL$ 농도의 들깨 추출물은 HCT116 대장암 세포와 H1299 폐암 세포의 성장을 18~94% 억제하는 농도 의존적 활성이 있었다. 또한 $175{\mu}g/mL$ 농도의 들깨 추출물은 HCT116과 H1299 세포에서 모두 colony 형성을 완전히 억제하는 활성이 있었다. 또한 들깨 추출물은 $87.5{\sim}350{\mu}g/mL$ 농도에서 H1299 세포의 이동을 30~37% 억제하였고, 가장 높은 농도인 $350{\mu}g/mL$ 농도에서는 HCT116과 H1299 세포의 부착을 14~16% 억제하였다. 이상의 결과를 통하여 들깨 추출물은 항산화 활성 및 암세포의 주요 특징을 억제하는 활성을 가지는 것으로 생각되며, 향후 이러한 연구 결과가 in vivo 수준에서 재현되는지 여부와 본 활성과 관련된 세부작용기전 또한 규명되어야 할 것으로 생각된다.

Keywords

References

  1. Lee CB. 2003. Coloured flora of Korea. 1st ed. Hyang Mun Sa, Seoul, Korea. p 140.
  2. Kim TJ. 1996. Korean resources plants IV. Seoul National University Publisher, Seoul, Korea. p 60.
  3. Ministry of Health & Welfare. 2010. Korea Health Statistics: Korea national health and nutrition examination survey (KNHANES) V-1. Ministry of Health & Welfare, Seoul, Korea. p 345-348.
  4. Park DS, Lee KI, Park KY. 2001. Antimutagenic effect of Perilla frutescens seed extract. J Korean Soc Food Sci Nutr 30: 900-905.
  5. Lee HA, Han JS. 2012. Anti-inflammatory effect of Perilla frutescens (L.) Britton var. frutescens extract in LPS-stimulated raw 264.7 macrophages. Prev Nutr Food Sci 17: 109-115. https://doi.org/10.3746/pnf.2012.17.2.109
  6. Lee Y, Song B, Ju J. 2014. Anti-inflammatory activity of perilla frutescens britton seed in RAW 264.7 macrophages and an ulcerative colitis mouse model. Korean J Food Sci Technol 46: 61-67. https://doi.org/10.9721/KJFST.2014.46.1.61
  7. Kim KH. 2003. Antioxidative effect of various solvent extracts from roasted perilla oil. Woosong Review 8: 1-11.
  8. Okuno M, Kajiwara K, Imai S, Kobayashi T, Honma N, Maki T, Suruga K, Goda T, Takase S, Muto Y, Moriwaki H. 1997. Perilla oil prevents the excessive growth of visceral adipose tissue in rats by down-regulating adipocyte differentiation. J Nutr 127: 1752-1757.
  9. Narisawa T, Takahashi M, Kotanagi H, Kusaka H, Yamazaki Y, Koyama H, Fukaura Y, Nishizawa Y, Kotsugai M, Isoda Y, Hirano J, Noritoshi N. 1991. Inhibitory effect of dietary perilla oil rich in the n-3 polyunsaturated fatty acid $\alpha$-linolenic acid on colon carcinogenesis in rats. Cancer Sci 82: 1089- 1096.
  10. Yoon SK, Kim JH, Kim ZU. 1993. Studies on antioxidant activity of ethanol extracts from defatted perilla flour. Korean J Food Sci Technol 25: 160-164.
  11. Cho HS, Ahn MS. 1999. Antioxidative effect of phenolic acids in defatted perilla flour on soybean oil. Korean J Food Cookery Sci 15: 55-60.
  12. Um MY, Choi WH, An JY, Kim SR, Ha TY. 2004. Effect of defatted sesame and perilla methanol extracts on cognitive function and antioxidant activity in SAMP8 mice. Korean J Food Sci Technol 36: 637-642.
  13. Rhie SG, Park YJ. 1991. Effect of the addition of defatted perilla on lipid metabolism in rats. J Korean Soc Food Sci Nutr 20: 72-77.
  14. National Academy of Agricultural Science. 2011. Food composition table. 8th ed. Rural Development Administration, Suwon, Korea. p 100.
  15. Paschos GK, Magkos F, Panagiotakos DB, Votteas V, Zampelas A. 2007. Dietary supplementation with flaxseed oil lowers blood pressure in dyslipidaemic patients. Eur J Clin Nutr 61: 1201-1206. https://doi.org/10.1038/sj.ejcn.1602631
  16. Holy EW, Forestier M, Richter EK, Akhmedov A, Leiber F, Camici GG, Mocharla P, Luscher TF, Beer JH, Tanner FC. 2011. Dietary $\alpha$-linolenic acid inhibits arterial thrombus formation, tissue factor expression, and platelet activation. Arterioscler Thromb Vasc Biol 31: 1772-1780. https://doi.org/10.1161/ATVBAHA.111.226118
  17. Djousse L, Pankow JS, Eckfeldt JH, Folsom AR, Hopkins PN, Province MA, Hong Y, Ellison RC. 2001. Relation between dietary linolenic acid and coronary artery disease in the National Heart, Lung, and Blood Institute Family Heart Study. Am J Clin Nutr 74: 612-619.
  18. Song JH, Park HS, Seo ES, Kim DY. 1994. Effect of different dietary fats on colon tumor incidence and in vivo cell proliferation in colonic mucosa of MNU-treated rats. Korean J Nutr 27: 552-562.
  19. Frankel EN, Huang SW, Aeschbach R, Prior E. 1996. Antioxidant activity of a rosemary extract and its constituents, carnosic acid, carnosol, and rosmarinic acid, in bulk oil and oil-in-water emulsion. J Agric Food Chem 44: 131-135. https://doi.org/10.1021/jf950374p
  20. Swarup V, Ghosh J, Ghosh S, Saxena A, Basu A. 2007. Antiviral and anti-inflammatory effects of rosmarinic acid in an experimental murine model of Japanese encephalitis. Antimicrob Agents Chemother 51: 3367-3370. https://doi.org/10.1128/AAC.00041-07
  21. Osakabe N, Yasuda A, Natsume M, Yoshikawa T. 2004. Rosmarinic acid inhibits epidermal inflammatory responses: anticarcinogenic effect of Perilla frutescens extract in the murine two-stage skin model. Carcinogenesis 25: 549-557.
  22. Osakabe N, Yasuda A, Natsume M, Sanbongi C, Kato Y, Osawa T, Yoshikawa T. 2002. Rosmarinic acid, a major polyphenolic component of Perilla frutescens, reduces lipopolysaccharide (LPS)-induced liver injury in D-galactosamine (D-GalN)-sensitized mice. Free Radic Biol Med 33: 798-806. https://doi.org/10.1016/S0891-5849(02)00970-X
  23. Furtado MA, de Almeida LC, Furtado RA, Cunha WR, Tavares DC. 2008. Antimutagenicity of rosmarinic acid in Swiss mice evaluated by the micronucleus assay. Mutat Res 657: 150-154. https://doi.org/10.1016/j.mrgentox.2008.09.003
  24. Chakraborty D, Mandal SM, Chakraborty J, Bhattacharyaa PK, Bandyopadhyay A, Mitra A, Gupta K. 2007. Antimicrobial activity of leaf extract of Basilicum polystachyon (L) Moench. Indian J Exp Biol 45: 774-748.
  25. Takano H, Osakabe N, Sanbongi C, Yanagisawa R, Inoue K, Yasuda A, Natsume M, Baba S, Ichiishi E, Yoshikawa T. 2004. Extract of Perilla frutescens enriched for rosmarinic acid, a polyphenolic phytochemical, inhibits seasonal allergic rhinoconjunctivitis in humans. Exp Biol Med 229: 247-254. https://doi.org/10.1177/153537020422900305
  26. Tsai SJ, Yin MC. 2008. Antioxidative and anti-inflammatory protection of oleanolic acid and ursolic acid in PC12 cells. J Food Sci 73: H174-H178. https://doi.org/10.1111/j.1750-3841.2008.00864.x
  27. Hsu HY, Yang JJ, Lin CC. 1997. Effects of oleanolic acid and ursolic acid on inhibiting tumor growth and enhancing the recovery of hematopoietic system postirradiation in mice. Cancer Lett 111: 7-13. https://doi.org/10.1016/S0304-3835(96)04481-3
  28. Sohn KH, Lee HY, Chung HY, Young HS, Yi SY, Kim KW. 1995. Anti-angiogenic activity of triterpene acids. Cancer Lett 94: 213-218. https://doi.org/10.1016/0304-3835(95)03856-R
  29. Valko M, Leibfritz D, Moncol J, Cronin MT, Mazur M, Telser J. 2007. Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol 39: 44-84. https://doi.org/10.1016/j.biocel.2006.07.001
  30. Kleinsmith LJ. 2008. Principles of cancer biology. 1st ed. Life Science Publishing Co., Seoul, Korea. p 16-63.
  31. World Health Organization. 2012. World health statistics 2012. World Health Organization, Geneva, Switzerland. p 80-81.
  32. Statistics Korea. 2012. 2011 Cause of death statistics. Statistics Korea, Daejeon, Korea. p 9-10.
  33. Peter B, Bernard L. 2008. World Cancer Report 2008. International Agency for Research on Cancer, Geneva, Switzerland. p 42-43.
  34. Ministry of Health and Welfare. 2012. Ministry of health and welfare statistical year book 2012. Ministry of Health & Welfare, Seoul, Korea. p 8-9.
  35. Singleton VL, Rossi Jr JA. 1965. Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. Am J Enol Viticult 16: 144-158.
  36. Zhishen J, Mengcheng T, Jianming W. 1999. The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chem 64: 555-559. https://doi.org/10.1016/S0308-8146(98)00102-2
  37. Brand-Williams W, Cuvelier ME, Berset C. 1995. Use of a free radical method to evaluate antioxidant activity. LWT-Food Sci Technol 28: 25-30. https://doi.org/10.1016/S0023-6438(95)80008-5
  38. Benzie IF, Strain JJ. 1996. The ferric reducing ability of plasma (FRAP) as a measure of "antioxidant power": the FRAP assay. Anal Biochem 239: 70-76. https://doi.org/10.1006/abio.1996.0292
  39. Lambert JD, Lu G, Lee MJ, Hu J, Ju J, Yang CS. 2009. Inhibition of lung cancer growth in mice by dietary mixed tocopherols. Mol Nutr Food Res 53: 1030-1035. https://doi.org/10.1002/mnfr.200800438
  40. Hou Z, Sang S, You H, Lee MJ, Hong J, Chin KV, Yang CS. 2005. Mechanism of action of (-)-epigallocatechin-3- gallate: auto-oxidation-dependent inactivation of epidermal growth factor receptor and direct effects on growth inhibition in human esophageal cancer KYSE 150 cells. Cancer Res 65: 8049-8056. https://doi.org/10.1158/0008-5472.CAN-05-0480
  41. Ju J, Kwak Y, Hao X, Yang CS. 2012. Inhibitory effects of calcium against intestinal cancer in human colon cancer cells and Apc (Min/+) mice. Nutr Res Pract 6: 396-404. https://doi.org/10.4162/nrp.2012.6.5.396
  42. Matsuura N, Miyamae Y, Yamane K, Nagao Y, Hamada Y, Kawaguchi N, Katsuki T, Hirata K, Sumi S, Ishikawa H. 2006. Aged garlic extract inhibits angiogenesis and proliferation of colorectal carcinoma cells. J Nutr 136: 842S- 846S.
  43. National Academy of Agricultural Science. 2009. Tables of food functional composition. 1st ed. Rural Development Administration, Suwon, Korea. p 201.
  44. Lee JH, Lee S. 1994 Analysis of phenolic substances content in Korean plant foods. Korean J Food Sci Technol 26: 310-316.
  45. Lee JH, Park KH, Lee MH, Kim HT, Seo WD, Kim JY, Baek IY, Jang DS, Ha TJ. 2013. Identification, characterisation, and quantification of phenolic compounds in the antioxidant activity-containing fraction from the seeds of Korean perilla (Perilla frutescens) cultivars. Food Chem 136: 843-852. https://doi.org/10.1016/j.foodchem.2012.08.057
  46. Hanahan D, Weinberg RA. 2011. Hallmarks of cancer: the next generation. Cell 144: 646-674. https://doi.org/10.1016/j.cell.2011.02.013
  47. Carvalho M, Ferreira PJ, Mendes VS, Silva R, Pereira JA, Jeronimo C, Silva BM. 2010. Human cancer cell antiproliferative and antioxidant activities of Juglans regia L. Food Chem Toxicol 48: 441-447. https://doi.org/10.1016/j.fct.2009.10.043
  48. Clarke MF, Dick JE, Dirks PB, Eaves CJ, Jamieson CH, Jones DL, Visvader J, Weissman IL, Wahl GM. 2006. Cancer stem cells-perspectives on current status and future directions: AACR Workshop on cancer stem cells. Cancer Res 66: 9339-9344. https://doi.org/10.1158/0008-5472.CAN-06-3126
  49. Kakarala M, Brenner DE, Korkaya H, Cheng C, Tazi K, Ginestier C, Liu S, Dontu G, Wicha MS. 2010. Targeting breast stem cells with the cancer preventive compounds curcumin and piperine. Breast Cancer Res Treat 122: 777-785. https://doi.org/10.1007/s10549-009-0612-x
  50. Srivastava RK, Tang SN, Zhu W, Meeker D, Shankar S. 2011. Sulforaphane synergizes with quercetin to inhibit self-renewal capacity of pancreatic cancer stem cells. Front Biosci 3: 515-528. https://doi.org/10.2741/e266
  51. Tang SN, Singh C, Nall D, Meeker D, Shankar S, Srivastava RK. 2010. The dietary bioflavonoid quercetin synergizes with epigallocathechin gallate (EGCG) to inhibit prostate cancer stem cell characteristics, invasion, migration and epithelial- mesenchymal transition. J Mol Signal 5: 14. https://doi.org/10.1186/1750-2187-5-14

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