Bracken-fern Extracts Induce Cell Cycle Arrest and Apoptosis in Certain Cancer Cell Lines

  • Roudsari, Motahhareh Tourchi (Department of Biology, Faculty of Sciences, Ferdowsi University of Mashhad) ;
  • Bahrami, Ahmad Reza (Department of Biology, Faculty of Sciences, Ferdowsi University of Mashhad) ;
  • Dehghani, Hesam (Embryonic and Stem Cell Research Group, Institute of Biotechnology, Ferdowsi University of Mashhad) ;
  • Iranshahi, Mehrdad (Biotechnology Research Center and School of Pharmacy, Mashhad University of Medical Sciences) ;
  • Matin, Maryam Moghadam (Department of Biology, Faculty of Sciences, Ferdowsi University of Mashhad) ;
  • Mahmoudi, Mahmud (Immunology Research Center of Bu Ali, Mashhad University of Medical Sciences)
  • Published : 2012.12.31


Bracken fern [Pteridium aquilinem (L.) kuhn (Dennstaedtiaceae)] is one of the most common species on the planet. It has been consumed by humans and animals for centuries. Use by some human groups is because they believe bracken fern is good for health as plant medicine. However, it is also one of the few known plants that can cause tumors in farm animals. Many interested groups have focused their attention on bracken fern because of these interesting features. In order to evaluate the biological effects of exposure to this plant in cellular level, human cancer cell lines were treated with the fern dichloromethane extracts and the genotoxic and cytotoxic effects were studied. Anti-proliferative/cytotoxic effects were evaluated by cell count, MTT assay and flow cytometry methods with three different cancer cell lines, TCC, NTERA2, and MCF-7, and two normal cells, HDF1 and HFF3. Pro-apoptotic effects of the extracts were determined by DAPI staining and comet assay, on TCC cancer cells compared to the normal control cell lines. Cellular morphology was examined by light microscopy. Our present study showed that the extract caused DNA damage and apoptosis at high concentrations ($200{\mu}g/mL$) and also it may induce cell cycle arrest (G2/M phase) at mild concentrations (50 and $30{\mu}g/mL$) depending on the cell type and tumor origin. These results indicate that bracken fern extract is a potent source of anticancer compounds that could be utilized pharmaceutically.


Bracken ferns;cell proliferation;apoptosis;cell cycle arrest


  1. Iranshahi M, Sahebkar A, Takasaki M, et al (2009). Cancer chemopreventive activity of the prenylated coumarin, umbelliprenin, in vivo. Eur J Cancer Prev, 18, 412-5.
  2. Lee KH (2004). Discovery and development of natural product-derived chemotherapeutic agents based on a medicinal chemistry approach. J Nat Prod, 73, 500-16.
  3. Li W, Lam MS, Birkeland A, et al (2006). Cell-based assays for profiling activity and safety properties of cancer drugs. J Pharmacol Toxicol Methods, 54, 313-9.
  4. Lugli E, Ferraresi R, Roat E, et al (2009). Quercetin inhibits lymphocyte activation and proliferation without inducing apoptosis in peripheral mononuclear cells. Leuk Res, 33, 140-50.
  5. Mosmann T (1983). Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assay. J Immunol, 65, 55-63.
  6. Pereira LO, Bicalho LS, Campos-da-paz Lopes M, et al (2009). DNA damage and apotosis induced by Pteridium aquilinum aqueos extract in the oral cell lines HSG and OSCC-3. J Oral Pathol Med, 38, 441-7.
  7. Sahebkar A, Iranshahi M (2010). Cancer chemoprevention by 7-prenyloxycoumarins: a role for 5-lipoxygenase inhibition? Integr Cancer Ther, 9, 259-60.
  8. Sardon D, de la Fuente I, Calonge E, et al (2005). H-ras immunohistochemical expression and molecular analysis of urinary bladder lesions in grazing adult cattle exposed to bracken fern. J Comp Pathol, 132, 195-201.
  9. Siman SE, Povey AC, Ward TH, et al (2000). Fern spore extracts can damage DNA. Br J Cancer, 83, 69-73.
  10. Singh NP, McCoy MT, Tice RR, et al (1988). A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res, 175, 184-91.
  11. Sugimura T (2000). Nutrition and dietarycarcinogens. Carcinogenesis, 21, 387-95.
  12. Talylor JA (1990). The bracken problem: a global perspective. AIAS Occas Publ, 40, 3-19.
  13. Wilson D, Donaldson LJ, Sepai O (1998). should we be frightened of bracken? A review of the evidence. J Epidemiol Community Health, 52, 812-7.
  14. Alonso-Amelot ME, Avendano M (2002). Human carcinogenesis and bracken fern: a review of the evidence. Curr Med Chem, 9, 675-6.
  15. Backorova M, Backor M, Mikes J, et al (2011). Variable responses of different human cancer cells to the lichen compounds parietin, atranorin, usnic acid and gyrophoric acid. Toxic In Vitro, 25, 37-44.
  16. Barthomeuf C, Lim S, Iranshahi M, et al (2008). Umbelliprenin from Ferula szowitsiana inhibits the growth of human M4Beu metastatic pigmented malignant melanoma cells through cell-cycle arrest in G1 and induction of caspase-dependent apoptosis. Phytomedicine, 15, 103-11.
  17. Bruggisser R, von Daeniken K, Jundt G, et al (2002). Interference of plant extracts, phytoestrogens and antioxidants with the MTT tetrazolium assay. Planta Med, 68, 445-8.
  18. Checchi PM, Nettles JH, Zhou J, et al (2003). Microtubule-interacting drugs for cancer treatment. Trends Pharmacol Sci, 24, 361-5.
  19. Freitas RN, O'Connor PJ, Prakash AS, et al (2001). Bracken (Pteridium aquilinum)-induced DNA adducts in mouse tissues are different from the adduct inducedby the activated form of the Bracken carcinogen ptaquiloside. Biochem Biophys Res Commun, 281, 589-94.
  20. Gibellini L, Pinti M, Nasi M, et al (2011). Quercetin and cancer cheoprevention. Evid Based Complement Alternat Med, 2011, 591356
  21. Gupta K, Panda D (2002). Perturbation of microtubule polymerization by quercetin through tubulin binding: a novel mechanism of its antiproliferative activity. Biochemistry, 41, 13029.
  22. Honore S, Pasquier E, Braguer D (2005). Understanding microtubule dynamics for improved cancer therapy. Cell Mol Life Sci, 62, 3039-56.
  23. Iranshahi M, Arfa P, Ramezani M, et al (2007). Sesquiterpene coumarins from Ferula szowitsiana and in vitro antileishmanial activity of 7-prenyloxycoumarins against promastigotes. Phytochemistry, 68, 554-61.
  24. Iranshahi M, Kalategi F, Rezaee R, et al (2008). Cancer chemopreventive activity of terpenoid coumarins from Ferula species. Planta Med, 74, 147-50.

Cited by

  1. Multiple Effects of Bracken Fern under in vivo and in vitro Conditions vol.15, pp.18, 2014,
  2. Bracken fern toxicity and its associated clinicopathological effects in humans and animals: a review pp.1618-565X, 2018,