3,5-Dihydroxy-4-isopropylstilbene is a natural phytoalexin and was first identified as bacterial secondary metabolites. The aim of this study is to investigate in vitro antioxidant and anticancer activity of 3,5-dihydroxy-4-isopropystilbene purified from the cell free culture filtrate of Bacillus sp. N strain associated with rhabditid entomopathogenic nematode. Antioxidant activity was evaluated by five separate methods: free radical scavenging, reducing power assay, chelating effects on ferrous ions, NBT superoxide radical scavenging assay and hydroxyl radical scavenging activity. The stilbene recorded powerful antioxidant activity at various antioxidant systems in vitro. The superoxide radical scavenging (92.1 %) and hydroxyl radical scavenging (83.4 %) activities of the stilbenes at 100 μg/ml were higher than the butylated hydroxyanisole, the known antioxidant agent. Anticancer activity of stilbene was tested against breast cancer (MDAM B-231), cervical cancer (HeLa), lung cancer (A 549), colon cancer (HTL 116) cell lines using MTT method. The induction of apoptosis was studied by morphological analysis, apoptotic cell staining, caspase 3 activation assay and cell cycle analysis using flow cytometry. Stilbene induced significant morphological changes and DNA fragmentation associated with apoptosis in HeLa cells. Acridine orange/ethidium bromide stained cells indicated apoptosis induction by stilbene. Up-regulation of caspase 3 activity was also found in cells treated with stilbene. Flow cytometry analysis showed an increase in the percentage of apoptotic cells in sub G0 phase (2.4 % in control plates to 11.4 % in 25 μg/ml of stilbene) confirming the stilbene induced apoptosis. The results of the present study showed that stilbene demonstrated a strong antioxidant and anticancer effects. These suggest that stilbene may be used as possible natural antioxidant and anticancer agents to control various human diseases.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Ahmad, A., F.A. Syed, S. Singh, and S.M. Hadi. 2005. Prooxidant activity of resveratrol in the presence of copper ions: Mutagenicity in plasmid DNA. Toxicology Letters 159: 1–12.
Alarco′n de la Lastra, C., and I. Villegas. 2007. Resveratrol as an antioxidant and pro-oxidant agent: Mechanisms and clinical implications. Biochemical Society Transactions 35: 1156–1160.
Anto, R.J., M. Venkataraman, and D. Karunagaran. 2003. Inhibition of NF-KB Sensitizes A431 cells to epidermal growth factor-induced apoptosis, whereas its activation by ectopic expression of RelA confers resistance. Journal of Biological Chemistry 28: 25490–25498.
Butler, M.S. 2005. Natural products to drugs: Natural product derived compounds in clinical trials. Natural Products Reports 22: 162–195.
Butler, M.S. 2008. Natural products to drugs: Natural product-derived compounds in clinical trials. Natural Products Reports 25: 475–516.
Carlsen, M.H., B.L. Halvorsen, K. Holteet, S.K. Bøhn, S. Dragland, L. Sampson, C. Willey, H. Senoo, Y. Umezono, C. Sanada, I. Barikmo, N. Berhe, W.C. Willett, K.M. Phillips, D.R. Jacobs, and R. Blomhof. 2010. The total antioxidant content of more than 3100 foods, beverages, spices, herbs and supplements used worldwide. Nutrition Journal 9: 1–11.
Chung, S.K., T. Osawa, and S. Kawakishi. 1997. Hydroxyl radical scavenging effects of species and scavengers from brown mustard (Brassica nigra). Bioscience Biotechnology and Biochemistry 61: 118–123.
Cohen, J.J. 1991. Programmed cell death in the immune system. Advances in Immunology 50: 55–85.
Cragg, G.M., D.G.I. Kingston, and D.J. Newman. 2005. Anticancer agents from natural products. Boca Raton: CRC Press.
Dinis, T.C.P., V.M.C. Madeira, and L.M. Almeida. 1994. Action of phenolic derivatives (acetaminophen, salicylate, and 5-aminosalicylate) as inhibitors of membrane lipid peroxidation and as peroxyl radical scavengers. Archives of Biochemistry and Biophysics 315: 161–169.
Finkel, T., and N.J. Holbrook. 2000. Oxidants, oxidative stress and the biology of aging. Nature 408(6809): 239–247.
Girotti, A.W. 1998. Lipid hydroperoxide generation, turnover, and effector action in biological systems. Journal of Lipid Research 39: 1529–1542.
Grice, H.C. 1986. Safety evaluation of butylated hydroxytoluene (BHT) in the liver, lung and gastrointestinal tract. Food and Chemical Toxicology 24: 1127–1130.
Gülcin, O., M. Kufrevio, S. Oktay, and M.E. Buyukokuro. 2004. Antioxidant, antimicrobial, antiulcer and analgesic activities of nettle (Urtica dioica L.). Journal of Ethnopharmacology 90: 205–215.
Hagiwara, K., K. Nobuyoshi, Y. Yoshioka, R. Takahashi, F. Takeshita, and T. Ochiya. 2012. Stilbene derivatives promote Ago2-dependent tumour-suppressive microRNA activity. Scientific Reports 314: 1–8.
Halliwell, B., R. Aeschbach, J. Loligeret, and O.I. Aruoma. 1995. The characterization of antioxidants. Food and Chemical Toxicology 33: 601–617.
Hu, K., and J.M. Webster. 2000. Antibiotic production in relation to bacterial growth and nematode development in Photorhabdus-Heterorhabditis infected Galleria mellonella larvae. FEMS Microbiology Letters 189: 219–223.
Hu, K.J., J.X. Li, and W.J. Wand. 1998. Comparison of metabolites produced in vitro and in vivo by Photorhabdus luminescens, a bacterial symbiont of the entomopathogenic nematode Heterorhabditis megidis. Canadian Journal of Microbiology 44: 1072–1077.
Kannan, R.R.R., R. Arumugam, and P. Anantharaman. 2010. In vitro antioxidant activities of ethanol extract from Enhalus acoroides (L.F.) Royle. Asian Pacific Journal of Tropical Medicine 3(11): 898–901.
Kerr, J.F., A.H. Wyllie, and A.R. Currie. 1972. Apoptosis: A basic biological phenomenon with wide-ranging implications in tissue kinetics. British Journal of Cancer 26(4): 239–257.
Kinsella, J.E., E. Frankel, B. German, and J. Kanner. 1993. Possible mechanism for the protective role of the antioxidant in wine and plant foods. Food Technology 47: 85–89.
Kohen, R., and A. Nyska. 2002. Oxidation of biological systems: Oxidative stress phenomena, antioxidants, redox reactions, and methods for their quantification. Toxicologic Pathology 30: 620–650.
Konzen, M., D. De Marco, C.A.S. Cordova, T.O. Vieira, R.V. Antonio, and T.B. Creczynski-Pasa. 2006. Antioxidant properties of violacein: Possible relation on its biological function. Bioorganic and Medicinal Chemistry 14: 8307–8313.
Kuate, D., B.C.O. Etoundi, Y.B. Soukontoua, J.L. Ngondi, and J.E. Oben. 2010. Antioxidant characteristics of Dichrostachys glomerata spice extracts. CyTA—Journal of Food 8: 23–37.
Kumar, R., D.D. Bansal, D.D. Patel, S. Mishra, Y. Karamalakova, A. Zheleva, V. Gadjeva, and R.K. Sharma. 2011. Antioxidative and radioprotective activities of semiquinone glucoside derivative (SQGD) isolated from Bacillus sp. INM-1. Molecular and Cellular Biochemistry 349(1–2): 57–67.
Kumar, S.N., J.V. Siji, B. Nambisan, K.N. Rajasekharan, and C. Mohandas. 2012. Bioactive stilbenes from a Bacillus sp. N strain associated with a novel rhabditid entomopathogenic nematode. Letters Applied Microbiology 54: 410–417.
Lai, L.S., S.T. Chou, and W.W. Chao. 2001. Studies on the antioxidative activities of Hsian-tsao (Mesona procumbens Hemsl) leaf gum. Journal of Agriculture and Food Chemistry 49: 963–968.
Lee, J.Y., W.I. Hwang, and S.T. Lim. 2004. Antioxidant and anticancer activities of extracts from Platycodon grandiflorum A. de Candolle roots. Journal of Ethnopharmacology 93: 409–415.
Li, J., G.H. Chen, H.M. Wu, and J.M. Webster. 1995. Identification of two pigments and a hydroxystilbene antibiotic from Photorhabdus luminescens. Applied and Environment Microbiology 61: 4329–4333.
Liu, F., and T.B. Ng. 2000. Antioxidative and free radical scavenging activities of selected medicinal herbs. Life Sciences 66: 725–735.
Liu, F., V.E.C. Ooi, and S.T. Chang. 1997. Free radical scavenging activities of mushroom polysaccharide extracts. Life Sciences 60: 763–771.
Liu, J., C. Wang, Z. Wang, C. Zhang, S. Lu, and J. Liu. 2011. The antioxidant and free-radical scavenging activities of extract and fractions from corn silk (Zea mays L.) and related flavone glycosides. Food Chemistry 126: 261–269.
Lockshin, R.A., and Z. Zakeri. 2002. Caspase-independent cell deaths. Current Opinion in Cell Biology 14(6): 727–733.
Nawar, W.F. 1996. Lipids. In Food chemistry, 3rd ed, ed. O. Fennema, 225–320. New York: Marcel Dekker Inc.
Newman, D.J., and G.M. Cragg. 2007. Natural product as source of new drugs over the last 25 years. Journal of Natural Products 70: 461–477.
Oktay, M., J. Gulcin, and O. Kufrevio. 2003. Determination of in vitro antioxidant activity of fennel (Foeniculum vulgare) seed extracts. LWT. Food Science and Technology 36: 263–271.
Oyaizu, M. 1986. Studies on product of browning reaction prepared from glucose amine. Japanese Journal of Nutrition 44: 307–315.
Richardson, W.H., T.M. Schmidt, and K.H. Nealson. 1988. Identification of an anthraquinone pigment and a hydroxystilbene antibiotic from Xenorhabdus luminescens. Applied and Environment Microbiology 54: 1602–1605.
Sherwin, E.R., A.L. Branen, P.M. Davidson, and S. Salminen. 1990. Food additives, 139–193. New York: Marvel Dekker Inc.
Tadhani, M.B., V.H. Patel, and R. Subhash. 2007. In vitro antioxidant activities of Stevia rebaudiana leaves and callus. Journal of Food Composition and Analysis 20: 323–329.
Tang, L.R., G.H. Chen, J. Li, and B. Webster. 2007. Anti-inflammatory activities of stilbene analogs for targeting autoimmune diseases. Clinical Immunology 123: S124.
Watanabe, M., and T.S. Todo. 2008. The novel antioxidant edaravone: From bench 435 to bedside. Cardiovascular Therapeutics 26: 101–114.
Wyllie, A.H. 1980. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 284: 555–556.
Yen, G.C., and H.Y. Chen. 1995. Antioxidant activity of various tea extracts in relation to their antimutagenicity. Journal of Agriculture and Food Chemistry 43: 27–32.
Yoshida, H., H. Yanai, Y. Namiki, K. Fukatsu-Sasaki, N. Furutani, and N. Tada. 2006. Neuroprotective effects of edaravone: A novel free radical scavenger in cerebrovascular injury. CNS Drug Reviews 438: 9–20.
Zhang, E.X., L.J. Yu, Y.L. Zhou, and X. Xiao. 1996. Studies on the peroxidation of polyunsaturated fatty acid from lipoprotein induced by iron and the evaluation of 444 the anti-oxidative activity of some natural products. Acta Biochimica et Biophysica Sinica 28: 218–222.
The authors are grateful to Indian Council Medical Research (ICMR), Government of India for funding. We thank the Director, CTCRI, for providing facilities for the work.
Conflict of interest
Authors declared no conflict of interest.
Electronic supplementary material
Below is the link to the electronic supplementary material.
About this article
Cite this article
Kumar, S.N., Nambisan, B., Kumar, B.S.D. et al. Antioxidant and anticancer activity of 3,5-dihydroxy-4-isopropylstilbene produced by Bacillus sp. N strain isolated from entomopathogenic nematode. Arch. Pharm. Res. (2013). https://doi.org/10.1007/s12272-013-0207-2