Antioxidant and Apoptotic Effects of an Aqueous Extract of Urtica dioica on the MCF-7 Human Breast Cancer Cell Line

  • Fattahi, Sadegh (Cellular and Molecular Biology Research Center, Babol University of Medical Sciences) ;
  • Ardekani, Ali Motevalizadeh (Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR) ;
  • Zabihi, Ebrahim (Cellular and Molecular Biology Research Center, Babol University of Medical Sciences) ;
  • Abedian, Zeinab (Cellular and Molecular Biology Research Center, Babol University of Medical Sciences) ;
  • Mostafazadeh, Amrollah (Cellular and Molecular Biology Research Center, Babol University of Medical Sciences) ;
  • Pourbagher, Roghayeh (Cellular and Molecular Biology Research Center, Babol University of Medical Sciences) ;
  • Akhavan-Niaki, Haleh (Cellular and Molecular Biology Research Center, Babol University of Medical Sciences)
  • Published : 2013.09.30


Breast cancer is the most prevalent cancer and one of the leading causes of death among women in the world. Plants and herbs may play an important role in complementary or alternative treatment. The aim of this study was to evaluate the antioxidant and anti-proliferative potential of Urtica dioica. The anti oxidant activity of an aqueous extract of Urtica dioica leaf was measured by MTT assay and the FRAP method while its anti-proliferative activity on the human breast cancer cell line (MCF-7) and fibroblasts isolated from foreskin tissue was evaluated using MTT assay. Mechanisms leading to apoptosis were also investigated at the molecular level by measuring the amount of anti and pro-apoptotic proteins and at the cellular level by studying DNA fragmentation and annexin V staining by flow cytometry. The aqueous extract of Urtica dioica showed antioxidant effects with a correlation coefficient of $r^2$=0.997. Dose-dependent and anti-proliferative effects of the extract were observed only on MCF-7 cells after 72 hrs with an $IC_{50}$ value of 2 mg/ml. This anti proliferative activity was associated with an increase of apoptosis as demonstrated by DNA fragmentation, the appearance of apoptotic cells in flow cytometry analysis and an increase of the amount of calpain 1, calpastatin, caspase 3, caspase 9, Bax and Bcl-2, all proteins involved in the apoptotic pathway. This is the first time such in vitro antiproliferative effect of aqueous extract of Urtica dioica leaf has been described for a breast cancer cell line. Our findings warrant further research on Urtica dioica as a potential chemotherapeutic agent for breast cancer.


Breast cancer;Urtica dioica;MCF-7;apoptosis;anti oxidant


  1. Abu-Dahab Rand Afifi F (2007). Antiproliferative activity of selected medicinal plants of Jordan against a breast adenocarcinoma cell line (MCF7). Scientia Pharmaceutica, 75, 121-36.
  2. Apak R, Guclu K, Demirata B, et al (2007). Comparative evaluation of various total antioxidant capacity assays applied to phenolic compounds with the CUPRAC assay. Molecules, 12, 1496-547.
  3. Benzie IF, Strain JJ (1996). The ferric reducing ability of plasma (FRAP) as a measure of "antioxidant power": the FRAP assay. Anal Biochem, 239, 70-6.
  4. Bnouham M, Merhfour FZ, Ziyyat A, et al (2010). Antidiabetic effect of some medicinal plants of Oriental Morocco in neonatal non-insulin-dependent diabetes mellitus rats. Hum Exp Toxicol, 29, 865-71.
  5. Bosetti C, Spertini L, Parpinel M, et al (2005). Flavonoids and breast cancer risk in Italy. Cancer Epidemiol Biomarkers Prev, 14, 805-8.
  6. Bradford MM (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem, 72, 248-54.
  7. Bray F, Jemal A, Grey N, et al (2012). Global cancer transitions according to the Human Development Index (2008-2030): a population-based study. Lancet Oncol, 13, 790-801.
  8. Campbell MK, Carr C, Devellis B, et al (2009). A randomized trial of tailoring and motivational interviewing to promote fruit and vegetable consumption for cancer prevention and control. Ann Behav Med, 38, 71-85.
  9. Chen J, Grant ME, Schor AM, et al (1989). Differences between adult and foetal fibroblasts in the regulation of hyaluronate synthesis: correlation with migratory activity. J Cell Sci, 94, 577-89.
  10. Cheng EH, Kirsch DG, Clem RJ, et al (1997). Conversion of Bcl-2 to a Bax-like death effector by caspases. Science, 278, 1966-8.
  11. Cui Q, Yu JH, Wu JN, et al (2007). P53-mediated cell cycle arrest and apoptosis through a caspase-3-independent, but caspase-9-dependent pathway in oridonin-treated MCF-7 human breast cancer cells. Acta Pharmacol Sin, 28, 1057-66.
  12. DeSantis C, Siegel R, Bandi P, et al (2011). Breast cancer statistics, 2011. CA Cancer J Clin, 61, 409-18.
  13. Devarajan E, Sahin AA, Chen JS, et al (2002). Down-regulation of caspase 3 in breast cancer: a possible mechanism for chemoresistance. Oncogene, 21, 8843-51.
  14. El Haouari M, Bnouham M, Bendahou M, et al (2006). Inhibition of rat platelet aggregation by Urtica dioica leaves extracts. Phytother Res, 20, 568-72.
  15. Guler ER (2013). Investigation of Chemopreventif Properties of Urtica dioica L., in MCF-7 and MDA 231 Breast Cancer Cell Lines. The New J Med, 30, 50-3.
  16. Harris MH, Thompson CB (2000). The role of the Bcl-2 family in the regulation of outer mitochondrial membrane permeability. Cell Death Differ, 7, 1182-91.
  17. Herrmann M, Lorenz HM, Voll R, et al (1994). A rapid and simple method for the isolation of apoptotic DNA fragments. Nucleic Acids Res, 22, 5506-7.
  18. Huang XJ, Ren W, Li J, et al (2013). Anti-inflammatory and anticancer activities of ethanol extract of pendulous monkshood root in vitro. Asian Pac J Cancer Prev, 14, 3569-73.
  19. Janicke RU (2009). MCF-7 breast carcinoma cells do not express caspase-3. Breast Cancer Res Treat, 117, 219-21.
  20. Janicke RU, Sprengart ML, Wati MR, Porter AG (1998). Caspase-3 is required for DNA fragmentation and morphological changes associated with apoptosis. J Biol Chem, 273, 9357-60.
  21. Konrad L, Muller HH, Lenz C, et al (2000). Antiproliferative effect on human prostate cancer cells by a stinging nettle root (Urtica dioica) extract. Planta Med, 66, 44-7.
  22. Lam L, Hu X, Aktary Z, et al (2009). Tamoxifen and ICI 182,780 increase Bcl-2 levels and inhibit growth of breast carcinoma cells by modulating PI3K/AKT, ERK and IGF- 1R pathways independent of ERalpha. Breast Cancer Res Treat, 118, 605-21.
  23. Liang Y, Yan C, Schor NF (2001). Apoptosis in the absence of caspase 3. Oncogene, 20, 6570-8.
  24. Lin B, Kolluri SK, Lin F, et al (2004). Conversion of Bcl-2 from protector to killer by interaction with nuclear orphan receptor Nur77/TR3. Cell, 116, 527-40.
  25. Mc Geea M, Hyland E, Campiani G, et al (2002). Caspase-3 is not essential for DNA fragmentation in MCF-7 cells during apoptosis induced by the pyrrolo-1, 5-benzoxazepine, PBOX-6 Febs. Lett, 515, 66-70.
  26. Muraina IA, Suleiman MM, Eloff JN (2009). Can MTT be used to quantify the antioxidant activity of plant extracts? Phytomedicine, 16, 665-8.
  27. Nijveldt RJ, van Nood E, van Hoorn DE, et al (2001). Flavonoids: a review of probable mechanisms of action and potential applications. Am J Clin Nutr, 74, 418-25.
  28. Nizamutdinova IT, Lee GW, Son KH et al (2008). Tanshinone I effectively induces apoptosis in estrogen receptor-positive (MCF-7) and estrogen receptor-negative (MDA-MB-231) breast cancer cells. Int J Oncol, 33, 485-91.
  29. O'Donovan N, Crown J, Stunell H, et al (2003). Caspase 3 in breast cancer. Clin Cancer Res, 9, 738-42.
  30. Ofir R, Seidman R, Rabinski T, et al (2002). Taxol-induced apoptosis in human SKOV3 ovarian and MCF7 breast carcinoma cells is caspase-3 and caspase-9 independent. Cell Death Differ, 9, 636-42.
  31. Olaku O,White JD (2011). Herbal therapy use by cancer patients: A literature review on case reports. Eur J Cancer, 47, 508-14.
  32. Oltvai ZN, Milliman CL, Korsmeyer SJ (1993). Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell, 74, 609-19.
  33. Pandamooz S, Hadipour A, Akhavan-Niaki H, et al (2012). Short exposure to collagenase and co-culture with mice embryonic pancreas improve the human dermal fibroblast culture. Biotechnol Appl Biochem, 59, 254-61.
  34. Peterson J, Dwyer J (1998). Flavonoids: dietary occurrence and biochemical activity. Nutr Res, 18, 1995-2018.
  35. Salehin D, Fromberg I, Haugk C, et al (2010). Immunhistochemical Analysis for Expression of Calpain 1, Calpain 2 and Calpastatin in Endometrial Cancer. Ant Can Res, 30, 2837-44.
  36. Schor SL, Schor AM, Rushton G (1988a). Fibroblasts from cancer patients display a mixture of both foetal and adult-like phenotypic characteristics. J Cell Sci, 90, 401-7.
  37. Schor SL, Schor AM, Howell A, et al (1987). Hypothesis: persistent expression of fetal phenotypic characteristics by fibroblasts is associated with an increased susceptibility to neoplastic disease. Exp Cell Biol, 55, 11-7.
  38. Schor SL, Schor AM, Grey AM, et al (1988b). Foetal and cancer patient fibroblasts produce an autocrine migrationstimulating factor not made by normal adult cells. J Cell Sci, 90, 391-9.
  39. Shim HY, Park JH, Paik HD, et al (2007). Acacetin-induced apoptosis of human breast cancer MCF-7 cells involves caspase cascade, mitochondria-mediated death signaling and SAPK/JNK1/2-c-Jun activation. Mol Cells, 24, 95-104.
  40. Suzuki N, Takimoto Y, Suzuki R, et al (2013). Efficacy of oral administration of lentinula eododes mycelia extract for breast cancer patients undergoing postoperative hormone therapy. Asian Pac J Cancer Prev, 14, 3469-72.
  41. Terry P, Terry JB, Wolk A (2001). Fruit and vegetable consumption in the prevention of cancer: an update. J Intern Med, 250, 280-90.
  42. Testai L, Chericoni S, Calderone V, et al (2002). Cardiovascular effects of Urtica dioica L. (Urticaceae) roots extracts: in vitro and in vivo pharmacological studies. J Ethnopharmacol, 81, 105-9.
  43. Tophkhane C, Yang S, Bales W, et al (2007). Bcl-2 overexpression sensitizes MCF-7 cells to genistein by multiple mechanisms. Int J Oncol, 31, 867-74.
  44. Yu MS, Lai CS, Ho YS, et al (2007). Characterization of the effects of anti-aging medicine Fructus lycii on beta-amyloid peptide neurotoxicity. Int J Mol Med, 20, 261-8.

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