DOI QR코드

DOI QR Code

Radish (Raphanus sativus L. leaf) ethanol extract inhibits protein and mRNA expression of $ErbB_2$ and $ErbB_3$ in MDA-MB-231 human breast cancer cells

  • Kim, Woo-Kyoung ;
  • Kim, Ji-Hae ;
  • Jeong, Da-Hee ;
  • Chun, Young-Hee ;
  • Kim, Sun-Hee ;
  • Cho, Kang-Jin ;
  • Chang, Moon-Jeong
  • Received : 2011.04.19
  • Accepted : 2011.06.13
  • Published : 2011.08.31

Abstract

In this study, we investigated the effects of the ethanol extract of aerial parts of Raphanus sativus L. (ERL) on breast cancer cell proliferation and gene expression associated with cell proliferation and apoptosis in MDA-MB-231 human breast cancer cells. The MDA-MB-231 cells were cultured in the presence or absence of various concentrations (100, 200, or 300 ${\mu}g$/mL) of ERL. ERL significantly decreased cell proliferation after 48 h of incubation (P < 0.05). The protein and mRNA expression of $ErbB_2$ were decreased significantly in a dose-dependent manner (P < 0.05). The protein expression of $ErbB_3$ was decreased significantly at an ERL concentration of 300 ${\mu}g$/mL (P < 0.05), and mRNA expression of $ErbB_3$ was decreased significantly in a dose-dependent manner (P < 0.05). The protein expression of Akt was decreased significantly at the ERL concentration of 200 ${\mu}g$/mL (P < 0.05), and the protein expression of pAkt was decreased significantly in a dose-dependent manner (P < 0.05). The mRNA expression of Akt was decreased significantly at the ERL concentration of 200 ${\mu}g$/mL ERL (P < 0.05). The protein and mRNA expression of Bax were increased significantly at ERL concentrations of 200 ${\mu}g$/mL or higher (P < 0.05). The protein expression of $Bcl_2$ was increased significantly at ERL concentrations of 100 ${\mu}g$/mL or higher (P < 0.05), and mRNA expression of $Bcl_2$ was increased significantly at an ERL concentration of 300 ${\mu}g$/mL (P < 0.05). In conclusion, we suggest that Raphanus sativus, L. inhibits cell proliferation via the ErbB-Akt pathway in MDA-MB-231 cells.

Keywords

Raphanus sativus L. ethanol extract;epidermal growth factor receptor;apoptosis;cell proliferation;MDA-MB-231 cell

References

  1. Ferlay J, Shin H, Bray F, Forman D, Mathers C, Parkin D. GLOBOCAN 2008, Cancer Incidence and Mortality Worldwide: IARC CancerBase No. 10. Lyon, France: International Agency for Research on Cancer; 2010.
  2. National Cancer Center [Internet]. Annual report of cancer statistics in Korea in 2008. [cited 2011 March 2]. Available from: http://ncc.re.kr/common/downloadByNTC.jsp?attnum=232&code=999_101.
  3. Korea Institute of Health and Social Affairs, Ministry of Health and Welfare Affairs [Internet]. Korea Health Statistics 2009, Korea National Health and Nutrition Examination Survey (KNHANES IV-3); [cited 2011 May 27]. Available from: http://knhanes.cdc.go.kr/.
  4. Kim BR, Park JH, Kim SH, Cho KJ, Chang MJ. Antihypertensive properties of dried radish leaves powder in spontaneously hypertensive rats. Korean J Nutr 2010;43:561-9. https://doi.org/10.4163/kjn.2010.43.6.561
  5. Beevi SS, Narasu ML, Gowda BB. Polyphenolics profile, antioxidant and radical scavenging activity of leaves and stem of Raphanus sativus L. Plant Foods Hum Nutr 2010;65:8-17. https://doi.org/10.1007/s11130-009-0148-6
  6. Scalbert A, Williamson G. Dietary intake and bioavailability of polyphenols. J Nutr 2000;130:2073S-2085S. https://doi.org/10.1093/jn/130.8.2073S
  7. Yang CS, Wang ZY. Tea and cancer. J Natl Cancer Inst 1993;85:1038-49. https://doi.org/10.1093/jnci/85.13.1038
  8. Seeram NP, Adams LS, Zhang Y, Lee R, Sand D, Scheuller HS, Heber D. Blackberry, black raspberry, blueberry, cranberry, red raspberry, and strawberry extracts inhibit growth and stimulate apoptosis of human cancer cells in vitro. J Agric Food Chem 2006;54:9329-39. https://doi.org/10.1021/jf061750g
  9. Pianetti S, Guo S, Kavanagh KT, Sonenshein GE. Green tea polyphenol epigallocatechin-3 gallate inhibits Her-2/neu signaling, proliferation, and transformed phenotype of breast cancer cells. Cancer Res 2002;62:652-5.
  10. Kameoka S, Leavitt P, Chang C, Kuo SM. Expression of antioxidant proteins in human intestinal Caco-2 cells treated with dietary flavonoids. Cancer Lett 1999;146:161-7. https://doi.org/10.1016/S0304-3835(99)00253-0
  11. Seshadri S, Nambiar VS. Kanjero (Digera arvensis) and drumstick leaves (Moringa oleifera): nutrient profile and potential for human consumption. Plants Hum Health Nutr Policy 2003;91:41-59.
  12. Papi A, Orlandi M, Bartolini G, Barillari J, Iori R, Paolini M, Ferroni F, Grazia Fumo M, Pedulli GF, Valgimigli L. Cytotoxic and antioxidant activity of 4-methylthio-3-butenyl isothiocyanate from Raphanus sativus L. (Kaiware Daikon) sprouts. J Agric Food Chem 2008;56:875-83. https://doi.org/10.1021/jf073123c
  13. Callahan R. Genetic alterations in primary breast cancer. Breast Cancer Res Treat 1989;13:191-203. https://doi.org/10.1007/BF02106570
  14. Slamon DJ, Clark GM. Amplification of c-erbB-2 and aggressive human breast tumors? Science 1988;240:1795-8. https://doi.org/10.1126/science.3289120
  15. Takeuchi K, Ito F. EGF receptor in relation to tumor development: molecular basis of responsiveness of cancer cells to EGFRtargeting tyrosine kinase inhibitors. FEBS J 2010;277:316-26. https://doi.org/10.1111/j.1742-4658.2009.07450.x
  16. Burgess AW, Cho HS, Eigenbrot C, Ferguson KM, Garrett TP, Leahy DJ, Lemmon MA, Sliwkowski MX, Ward CW, Yokoyama S. An open-and-shut case? Recent insights into the activation of EGF/ErbB receptors. Mol Cell 2003;12:541-52. https://doi.org/10.1016/S1097-2765(03)00350-2
  17. Citri A, Yarden Y. EGF-ERBB signalling: towards the systems level. Nat Rev Mol Cell Biol 2006;7:505-16. https://doi.org/10.1038/nrm1962
  18. Slamon DJ, Godolphin W, Jones LA, Holt JA, Wong SG, Keith DE, Levin WJ, Stuart SG, Udove J, Ullrich A. Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer. Science 1989;244:707-12. https://doi.org/10.1126/science.2470152
  19. Bold RJ, Termuhlen PM, McConkey DJ. Apoptosis, cancer and cancer therapy. Surg Oncol 1997;6:133-42. https://doi.org/10.1016/S0960-7404(97)00015-7
  20. Sen P, Mukherjee S, Ray D, Raha S. Involvement of the Akt/PKB signaling pathway with disease processes. Mol Cell Biochem 2003;253:241-6 https://doi.org/10.1023/A:1026020101379
  21. Slamon DJ, Clark GM, Wong SG, Levin WJ, Ullrich A, McGuire WL. Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science 1987;235:177-82. https://doi.org/10.1126/science.3798106
  22. Cooke T, Reeves J, Lannigan A, Stanton P. The value of the human epidermal growth factor receptor-2 (HER2) as a prognostic marker. Eur J Cancer 2001;37:3-10.
  23. Lewis S, Locker A, Todd JH, Bell JA, Nicholson R, Elston CW, Blamey RW, Ellis IO. Expression of epidermal growth factor receptor in breast carcinoma. J Clin Pathol 1990;43:385-9. https://doi.org/10.1136/jcp.43.5.385
  24. Mendelsohn J, Baselga J. Status of epidermal growth factor receptor antagonists in the biology and treatment of cancer. J Clin Oncol 2003;21:2787-99. https://doi.org/10.1200/JCO.2003.01.504
  25. Pao W, Miller V, Zakowski M, Doherty J, Politi K, Sarkaria I, Singh B, Heelan R, Rusch V, Fulton L, Mardis E, Kupfer D, Wilson R, Kris M, Varmus H. EGF receptor gene mutations are common in lung cancers from "never smokers" and are associated with sensitivity of tumors to gefitinib and erlotinib. Proc Natl Acad Sci U S A 2004;101:13306-11. https://doi.org/10.1073/pnas.0405220101
  26. Janmaat ML, Kruyt FA, Rodriguez JA, Giaccone G. Response to epidermal growth factor receptor inhibitors in non-small cell lung cancer cells: limited antiproliferative effects and absence of apoptosis associated with persistent activity of extracellular signal-regulated kinase or Akt kinase pathways. Clin Cancer Res 2003;9:2316-26.
  27. Yuan ZQ, Sun M, Feldman RI, Wang G, Ma X, Jiang C, Coppola D, Nicosia SV, Cheng JQ. Frequent activation of AKT2 and induction of apoptosis by inhibition of phosphoinositide-3-OH kinase/Akt pathway in human ovarian cancer. Oncogene 2000;19:2324-30. https://doi.org/10.1038/sj.onc.1203598
  28. Bellacosa A, de Feo D, Godwin AK, Bell DW, Cheng JQ, Altomare DA, Wan M, Dubeau L, Scambia G, Masciullo V, Ferrandina G, Benedetti Panici P, Mancuso S, Neri G, Testa JR. Molecular alterations of the AKT2 oncogene in ovarian and breast carcinomas. Int J Cancer 1995;64:280-5. https://doi.org/10.1002/ijc.2910640412
  29. Cardone MH, Roy N, Stennicke HR, Salvesen GS, Franke TF, Stanbridge E, Frisch S, Reed JC. Regulation of cell death protease caspase-9 by phosphorylation. Science 1998;282:1318-21. https://doi.org/10.1126/science.282.5392.1318
  30. Brunet A, Bonni A, Zigmond MJ, Lin MZ, Juo P, Hu LS, Anderson MJ, Arden KC, Blenis J, Greenberg ME. Akt promotes cell survival by phosphorylating and inhibiting a Forkhead transcription factor. Cell 1999;96:857-68. https://doi.org/10.1016/S0092-8674(00)80595-4
  31. Tang ED, Nuñez G, Barr FG, Guan KL. Negative regulation of the forkhead transcription factor FKHR by Akt. J Biol Chem 1999;274:16741-6. https://doi.org/10.1074/jbc.274.24.16741
  32. Chun KH, Kosmeder JW 2nd, Sun S, Pezzuto JM, Lotan R, Hong WK, Lee HY. Effects of deguelin on the phosphatidylinositol 3-kinase/Akt pathway and apoptosis in premalignant human bronchial epithelial cells. J Natl Cancer Inst 2003;95:291-302. https://doi.org/10.1093/jnci/95.4.291
  33. Ow YP, Green DR, Hao Z, Mak TW. Cytochrome c: functions beyond respiration. Nat Rev Mol Cell Biol 2008;9:532-42. https://doi.org/10.1038/nrm2434
  34. Binder C, Marx D, Binder L, Schauer A, Hiddemann W. Expression of Bax in relation to Bcl-2 and other predictive parameters in breast cancer. Ann Oncol 1996;7:129-33. https://doi.org/10.1093/oxfordjournals.annonc.a010538
  35. Lee HS, Kim EJ, Kim SH. Chestnut extract induces apoptosis in AGS human gastric cancer cells. Nutr Res Pract 2011;5:185-91. https://doi.org/10.4162/nrp.2011.5.3.185
  36. Sakakura C, Sweeney EA, Shirahama T, Igarashi Y, Hakomori S, Nakatani H, Tsujimoto H, Imanishi T, Ohgaki M, Ohyama T, Yamazaki J, Hagiwara A, Yamaguchi T, Sawai K, Takahashi T. Overexpression of bax sensitizes human breast cancer MCF-7 cells to radiation-induced apoptosis. Int J Cancer 1996;67:101-5. https://doi.org/10.1002/(SICI)1097-0215(19960703)67:1<101::AID-IJC17>3.0.CO;2-H
  37. Wolter KG, Hsu YT, Smith CL, Nechushtan A, Xi XG, Youle RJ. Movement of Bax from the cytosol to mitochondria during apoptosis. J Cell Biol 1997;139:1281-92. https://doi.org/10.1083/jcb.139.5.1281
  38. Tsuruta F, Masuyama N, Gotoh Y. The phosphatidylinositol 3-kinase (PI3K)-Akt pathway suppresses Bax translocation to mitochondria. J Biol Chem 2002;277:14040-7. https://doi.org/10.1074/jbc.M108975200
  39. Korsmeyer SJ, Wei MC, Saito M, Weiler S, Oh KJ, Schlesinger PH. Pro-apoptotic cascade activates BID, which oligomerizes BAK or BAX into pores that result in the release of cytochrome c. Cell Death Differ 2000;7:1166-73. https://doi.org/10.1038/sj.cdd.4400783
  40. Shimizu S, Narita M, Tsujimoto Y. Bcl-2 family proteins regulate the release of apoptogenic cytochrome c by the mitochondrial channel VDAC. Nature 1999;399:483-7. https://doi.org/10.1038/20959

Cited by

  1. The effects of Gamijinhae-tang on elastase/lipopolysaccharide-induced lung inflammation in an animal model of acute lung injury vol.13, pp.1, 2013, https://doi.org/10.1186/1472-6882-13-176
  2. RsERF1 derived from wild radish (Raphanus sativus) confers salt stress tolerance in Arabidopsis vol.36, pp.4, 2014, https://doi.org/10.1007/s11738-013-1478-4
  3. Estonian folk traditional experiences on natural anticancer remedies: From past to the future vol.52, pp.7, 2014, https://doi.org/10.3109/13880209.2013.871641
  4. Biological activity of various radish species vol.15, pp.2, 2015, https://doi.org/10.1007/s13596-015-0183-9
  5. A New AP2/ERF Transcription Factor from the Oil Plant Jatropha curcas Confers Salt and Drought Tolerance to Transgenic Tobacco vol.176, pp.2, 2015, https://doi.org/10.1007/s12010-015-1597-z
  6. Quality Characteristics of Seolgiddeok with Radish (Raphanus sativus L.) Leaf Powder vol.31, pp.3, 2015, https://doi.org/10.9724/kfcs.2015.31.3.318
  7. Quality Characteristics of Soy Sauce Braised Shiraegi with Baknig Powder vol.31, pp.5, 2015, https://doi.org/10.9724/kfcs.2015.31.5.676
  8. Green synthesis of biogenic silver nanomaterials using Raphanus sativus extract, effects of stabilizers on the morphology, and their antimicrobial activities vol.38, pp.12, 2015, https://doi.org/10.1007/s00449-015-1477-5
  9. Effects of (6)-gingerol, ginger component on adipocyte development and differentiation in 3T3-L1 vol.48, pp.4, 2015, https://doi.org/10.4163/jnh.2015.48.4.327
  10. Toxicity Comparison of Four Cruciferous Plant Extracts and Evaluation of Their Cytotoxicity - Radical Scavenging Correlations vol.13, pp.2, 2018, https://doi.org/10.5812/jjnpp.13866
  11. Deciphering the Nutraceutical Potential of Raphanus sativus—A Comprehensive Overview vol.11, pp.2, 2019, https://doi.org/10.3390/nu11020402

Acknowledgement

Supported by : Rural Development Administration