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Inhibitory activities of Perilla frutescens britton leaf extract against the growth, migration, and adhesion of human cancer cells

  • Kwak, Youngeun (Department of Food and Nutrition, Chungbuk National University) ;
  • Ju, Jihyeung (Department of Food and Nutrition, Chungbuk National University)
  • Received : 2014.06.19
  • Accepted : 2014.10.31
  • Published : 2015.02.01

Abstract

BACKGROUND/OBJECTIVES: Perilla frutescens Britton leaves are a commonly consumed vegetable in different Asian countries including Korea. Cancer is a major cause of human death worldwide. The aim of the current study was to investigate the inhibitory effects of ethanol extract of perilla leaf (PLE) against important characteristics of cancer cells, including unrestricted growth, resisted apoptosis, and activated metastasis, using human cancer cells. MATERIALS/METHODS: Two human cancer cell lines were used in this study, HCT116 colorectal carcinoma cells and H1299 non-small cell lung carcinoma cells. Assays using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide were performed for measurement of cell growth. Soft agar and wound healing assays were performed to determine colony formation and cell migration, respectively. Nuclear staining and cell cycle analysis were performed for assessment of apoptosis. Fibronectin-coated plates were used to determine cell adhesion. RESULTS: Treatment of HCT116 and H1299 cells with PLE resulted in dose-dependent inhibition of growth by 52-92% (at the concentrations of 87.5, 175, and $350{\mu}g/ml$) and completely abolished the colony formation in soft agar (at the concentration of $350{\mu}g/ml$). Treatment with PLE at the $350{\mu}g/ml$ concentration resulted in change of the nucleus morphology and significantly increased sub-G1 cell population in both cells, indicating its apoptosis-inducing activity. PLE at the concentration range of 87.5 to $350{\mu}g/ml$ was also effective in inhibiting the migration of H1299 cells (by 52-58%) and adhesion of both HCT116 and H1299 cells (by 25-46%). CONCLUSIONS: These results indicate that PLE exerts anti-cancer activities against colon and lung cancers in vitro. Further studies are needed in order to determine whether similar effects are reproduced in vivo.

Acknowledgement

Supported by : National Research Foundation of Korea

References

  1. Jemal A, Siegel R, Ward E, Hao Y, Xu J, Thun MJ. Cancer statistics, 2009. CA Cancer J Clin 2009;59:225-49. https://doi.org/10.3322/caac.20006
  2. Surh YJ. Cancer chemoprevention with dietary phytochemicals. Nat Rev Cancer 2003;3:768-80. https://doi.org/10.1038/nrc1189
  3. Miller PE, Snyder DC. Phytochemicals and cancer risk: a review of the epidemiological evidence. Nutr Clin Pract 2012;27:599-612. https://doi.org/10.1177/0884533612456043
  4. Seo WH, Baek HH. Characteristic aroma-active compounds of Korean perilla (Perilla frutescens Britton) leaf. J Agric Food Chem 2009;57:11537-42. https://doi.org/10.1021/jf902669d
  5. Meng L, Lozano YF, Gaydou EM, Li B. Antioxidant activities of polyphenols extracted from Perilla frutescens varieties. Molecules 2008;14:133-40. https://doi.org/10.3390/molecules14010133
  6. Ueda H, Yamazaki M. Inhibition of tumor necrosis factor-alpha production by orally administering a perilla leaf extract. Biosci Biotechnol Biochem 1997;61:1292-5. https://doi.org/10.1271/bbb.61.1292
  7. Ueda H, Yamazaki M. Anti-inflammatory and anti-allergic actions by oral administration of a perilla leaf extract in mice. Biosci Biotechnol Biochem 2001;65:1673-5. https://doi.org/10.1271/bbb.65.1673
  8. Kim MK, Lee HS, Kim EJ, Won NH, Chi YM, Kim BC, Lee KW. Protective effect of aqueous extract of Perilla frutescens on tert-butyl hydroperoxide-induced oxidative hepatotoxicity in rats. Food Chem Toxicol 2007;45:1738-44. https://doi.org/10.1016/j.fct.2007.03.009
  9. Makino T, Furuta A, Fujii H, Nakagawa T, Wakushima H, Saito K, Kano Y. Effect of oral treatment of Perilla frutescens and its constituents on type-I allergy in mice. Biol Pharm Bull 2001;24:1206-9. https://doi.org/10.1248/bpb.24.1206
  10. Liu JY, Chen YC, Lin CH, Kao SH. Perilla frutescens leaf extract inhibits mite major allergen Der p 2-induced gene expression of pro-allergic and pro-inflammatory cytokines in human bronchial epithelial cell BEAS-2B. PLoS One 2013;8:e77458. https://doi.org/10.1371/journal.pone.0077458
  11. Makino T, Furuta Y, Wakushima H, Fujii H, Saito K, Kano Y. Antiallergic effect of Perilla frutescens and its active constituents. Phytother Res 2003;17:240-3. https://doi.org/10.1002/ptr.1115
  12. Kim MJ, Kim HK. Perilla leaf extract ameliorates obesity and dyslipidemia induced by high-fat diet. Phytother Res 2009;23: 1685-90. https://doi.org/10.1002/ptr.2811
  13. Kwak CS, Yeo EJ, Moon SC, Kim YW, Ahn HJ, Park SC. Perilla leaf, Perilla frutescens, induces apoptosis and G1 phase arrest in human leukemia HL-60 cells through the combinations of death receptormediated, mitochondrial, and endoplasmic reticulum stress-induced pathways. J Med Food 2009;12:508-17. https://doi.org/10.1089/jmf.2008.1103
  14. Lin CS, Kuo CL, Wang JP, Cheng JS, Huang ZW, Chen CF. Growth inhibitory and apoptosis inducing effect of Perilla frutescens extract on human hepatoma HepG2 cells. J Ethnopharmacol 2007;112: 557-67. https://doi.org/10.1016/j.jep.2007.05.008
  15. Ueda H, Yamazaki C, Yamazaki M. Inhibitory effect of Perilla leaf extract and luteolin on mouse skin tumor promotion. Biol Pharm Bull 2003;26:560-3. https://doi.org/10.1248/bpb.26.560
  16. Jeon IH, Kim HS, Kang HJ, Lee HS, Jeong SI, Kim SJ, Jang SI. Anti-inflammatory and antipruritic effects of luteolin from Perilla (P. frutescens L.) leaves. Molecules 2014;19:6941-51. https://doi.org/10.3390/molecules19066941
  17. Ueda H, Yamazaki C, Yamazaki M. Luteolin as an anti-inflammatory and anti-allergic constituent of Perilla frutescens. Biol Pharm Bull 2002;25:1197-202. https://doi.org/10.1248/bpb.25.1197
  18. Osakabe N, Yasuda A, Natsume M, Sanbongi C, Kato Y, Osawa T, Yoshikawa T. 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 2002;33:798-806. https://doi.org/10.1016/S0891-5849(02)00970-X
  19. Osakabe N, Yasuda A, Natsume M, Yoshikawa T. Rosmarinic acid inhibits epidermal inflammatory responses: anticarcinogenic effect of Perilla frutescens extract in the murine two-stage skin model. Carcinogenesis 2004;25:549-57.
  20. Sanbongi C, Takano H, Osakabe N, Sasa N, Natsume M, Yanagisawa R, Inoue KI, Sadakane K, Ichinose T, Yoshikawa T. Rosmarinic acid in perilla extract inhibits allergic inflammation induced by mite allergen, in a mouse model. Clin Exp Allergy 2004;34:971-7. https://doi.org/10.1111/j.1365-2222.2004.01979.x
  21. Takano H, Osakabe N, Sanbongi C, Yanagisawa R, Inoue K, Yasuda A, Natsume M, Baba S, Ichiishi E, Yoshikawa T. Extract of Perilla frutescens enriched for rosmarinic acid, a polyphenolic phytochemical, inhibits seasonal allergic rhinoconjunctivitis in humans. Exp Biol Med (Maywood) 2004;229:247-54. https://doi.org/10.1177/153537020422900305
  22. Banno N, Akihisa T, Tokuda H, Yasukawa K, Higashihara H, Ukiya M, Watanabe K, Kimura Y, Hasegawa J, Nishino H. Triterpene acids from the leaves of Perilla frutescens and their anti-inflammatory and antitumor-promoting effects. Biosci Biotechnol Biochem 2004; 68:85-90. https://doi.org/10.1271/bbb.68.85
  23. Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell 2011;144:646-74. https://doi.org/10.1016/j.cell.2011.02.013
  24. Lambert JD, Lu G, Lee MJ, Hu J, Ju J, Yang CS. Inhibition of lung cancer growth in mice by dietary mixed tocopherols. Mol Nutr Food Res 2009;53:1030-5. https://doi.org/10.1002/mnfr.200800438
  25. Irons R, Tsuji PA, Carlson BA, Ouyang P, Yoo MH, Xu XM, Hatfield DL, Gladyshev VN, Davis CD. Deficiency in the 15-kDa selenoprotein inhibits tumorigenicity and metastasis of colon cancer cells. Cancer Prev Res (Phila) 2010;3:630-9. https://doi.org/10.1158/1940-6207.CAPR-10-0003
  26. Toton E, Ignatowicz E, Bernard MK, Kujawski J, Rybczynska M. Evaluation of apoptotic activity of new condensed pyrazole derivatives. J Physiol Pharmacol 2013;64:115-23.
  27. Fesik SW. Promoting apoptosis as a strategy for cancer drug discovery. Nat Rev Cancer 2005;5:876-85. https://doi.org/10.1038/nrc1736
  28. Florent M, Godard T, Ballet JJ, Gauduchon P, Sola B. Detection by the comet assay of apoptosis induced in lymphoid cell lines after growth factor deprivation. Cell Biol Toxicol 1999;15:185-92. https://doi.org/10.1023/A:1007641821779
  29. Ju J, Kwak Y, Hao X, Yang CS. Inhibitory effects of calcium against intestinal cancer in human colon cancer cells and Apc(Min/+) mice. Nutr Res Pract 2012;6:396-404. https://doi.org/10.4162/nrp.2012.6.5.396
  30. Matsuura N, Miyamae Y, Yamane K, Nagao Y, Hamada Y, Kawaguchi N, Katsuki T, Hirata K, Sumi S, Ishikawa H. Aged garlic extract inhibits angiogenesis and proliferation of colorectal carcinoma cells. J Nutr 2006;136:842S-846S. https://doi.org/10.1093/jn/136.3.842S
  31. Hotz MA, Gong J, Traganos F, Darzynkiewicz Z. Flow cytometric detection of apoptosis: comparison of the assays of in situ DNA degradation and chromatin changes. Cytometry 1994;15:237-44. https://doi.org/10.1002/cyto.990150309
  32. Li K, Zhu ZC, Liu YJ, Liu JW, Wang HT, Xiong ZQ, Shen X, Hu ZL, Zheng J. ZFX knockdown inhibits growth and migration of non-small cell lung carcinoma cell line H1299. Int J Clin Exp Pathol 2013;6:2460-7.
  33. Tang J, Zhang L, She X, Zhou G, Yu F, Xiang J, Li G. Inhibiting CD164 expression in colon cancer cell line HCT116 leads to reduced cancer cell proliferation, mobility, and metastasis in vitro and in vivo. Cancer Invest 2012;30:380-9. https://doi.org/10.3109/07357907.2012.666692
  34. Dolfi SC, Yang Z, Lee MJ, Guan F, Hong J, Yang CS. Inhibitory effects of different forms of tocopherols, tocopherol phosphates, and tocopherol quinones on growth of colon cancer cells. J Agric Food Chem 2013;61:8533-40. https://doi.org/10.1021/jf401076g
  35. Wang X, Wang Q, Ives KL, Evers BM. Curcumin inhibits neurotensin-mediated interleukin-8 production and migration of HCT116 human colon cancer cells. Clin Cancer Res 2006;12:5346-55. https://doi.org/10.1158/1078-0432.CCR-06-0968
  36. Lu G, Xiao H, Li GX, Picinich SC, Chen YK, Liu A, Lee MJ, Loy S, Yang CS. A gamma-tocopherol-rich mixture of tocopherols inhibits chemically induced lung tumorigenesis in A/J mice and xenograft tumor growth. Carcinogenesis 2010;31:687-94. https://doi.org/10.1093/carcin/bgp332
  37. Ballet F, Petit J, Poupon R, Darnis F. Soft agar clonogenic assay for predicting chemosensitivity of human tumor cells from malignant effusions. Biomedicine 1981;35:177-8.
  38. Shu L, Cheung KL, Khor TO, Chen C, Kong AN. Phytochemicals: cancer chemoprevention and suppression of tumor onset and metastasis. Cancer Metastasis Rev 2010;29:483-502. https://doi.org/10.1007/s10555-010-9239-y
  39. Lin Y, Shi R, Wang X, Shen HM. Luteolin, a flavonoid with potential for cancer prevention and therapy. Curr Cancer Drug Targets 2008;8:634-46. https://doi.org/10.2174/156800908786241050
  40. Wang Y, Huang X, Han J, Zheng W, Ma W. Extract of Perilla frutescens inhibits tumor proliferation of HCC via PI3K/AKT signal pathway. Afr J Tradit Complement Altern Med 2012;10:251-7.

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