Comparative analysis of the cytotoxic effect of 7-prenyloxycoumarin compounds and herniarin on MCF-7 cell line

Document Type : Original Research Article


1 Pharmacological Research Centre of Medicinal Plants, School of Medicine, Mashhad, University of Medical Sciences, Mashhad, Iran.

2 Medical Toxicology Research Centre, Mashhad, University of Medical Sciences, Mashhad, Iran.

3 Biotechnology Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.

4 Department of Pharmacodynamics and Toxicology, School of Pharmacy; Mashhad University of Medical Sciences, Mashhad, Iran.


Objectives: 7-prenyloxycoumarins are a group of secondary metabolites that are found mainly in plants belonging to the Rutaceae and Umbelliferae families. This study was designed to evaluate and compare the cytotoxic and apoptotic activity of 7-prenyloxycoumarin compounds and herniarin on MCF-7, a breast carcinoma cell line.
Materials and Methods: Cells were cultured in RPMI medium and incubated with different concentrations of auraptene, herniarin, umbelliferone, and umbelliprenin. Cell viability was quantified by MTT assay. Apoptotic cells were determined using propidium iodide staining of DNA fragmentation by flow cytometry (sub-G1peak). Bax protein expression was detected by western blot to investigate the underlying mechanism.
Results: Doses which induced 50% cell growth inhibition (IC50) against MCF-7 cells with auraptene, herniarin, umbelliferone, and umbelliprenin were calculated (59.7, 207.6, 476.3, and 73.4 µM), respectively. Auraptene induced a sub-G1 peak in the flow cytometry histogram of treated cells compared to control cells, and DNA fragmentation suggested the induction of apoptosis. Western blot analysis showed that auraptene significantly up-regulated Bax expression in MCF-7 cells compared to untreated controls.
Conclusion: Auraptene exerts cytotoxic and apoptotic effects in breast carcinoma cell line and can be considered for further mechanistic evaluations in human cancer cells. These results candidate auraptene for further studies to evaluate its biosafety and anti-cancer effects.


Main Subjects

Akins RE, Tuan RS. 1992. Measurement of protein in 20 seconds using a microwave BCA assay. BioTechniques, 12: 96-497.
Askari M, Sahebkar AH, Iranshahi M. 2009. Synthesis and Purification of 7 Prenyloxycoumarins and Herniarin as Bioactive Natural Coumarins. Iran J Basic Med Sci,12: 63-69.
Baba M, Jin Y, Mizuno A, Suzuki H, Okada Y, et al. 2002. Studies on cancer chemoprevention by traditional folk medicines XXIV Inhibitory effect of a coumarin derivative 7-isopentenyloxycoumarin, against tumor-promotion. Biol Pharm Bull, 25: 244-246.
Barthomeuf C, Lim S, Iranshahi M, Chollet P. 2008. Umbelliprenin from Ferula szowitsiana inhibits the growth of human M4Beu metastatic pigmented melanoma cells through cell-cycle arrest in G1 and induction of caspasedependent apoptosis. Phytomedicine, 15: 103-111.
Cassileth B, Yeung KS, Gubili J. 2008. Herbs and Other Botanicals in Cancer Patient Care. Curr Treat Options, Oncol, 9: 109-16.
Chen IS, Lin YC, Tsai IL, Teng CM, Ko FN, et al. 1995. Coumarins and anti-platelet aggregation constituents from Zanthoxylum schinifolium. Phytochemistry, 39: 1091-1097.
Curini M, Epifano F, Maltese F, Marcotullioa MC, Tubarob A, et al. 2004. Synthesis and anti-inflammatory activity of natural and semisynthetic geranyloxycoumarins. Bioorg Med Chem Lett,14: 2241-2243.
de Medina P, Genovese S, Paillasse MR, Mazaheri M, Caze-Subra S, Bystricky K, Curini M, Silvente-Poirot S, Epifano F, Poirot M. 2010. Auraptene is an inhibitor of cholesterol esterification and a modulator of estrogen receptors. Mol Pharmacol, 78: 827-36.
Gong J, Traganos F, Darzynkiewicz Z. 1994. A selective procedure for DNA extraction from apoptotic cells applicable for gel electrophoresis and flow cytometry. Anal Biochem, 218: 314-319.
Gross A, McDonnell JM, Korsmeyer SJ. 1999. BCL-2 family members and the mitochondria in apoptosis. Genes Dev, 13: 1899-1911.
Iranshahi M, Arfa P, Ramezani M, Jaafari MR, Sadeghian H, et al. 2007. Sesquiterpene coumarins from Ferula szowitsiana and in vitro antileishmanial activity of 7-prenyloxycoumarins against promastigotes, Phytochemistry, 68: 554-561.
Iranshahi M, Kalategi F, Rezaee R, Shahverdi AR, Ito C, et al. 2008. Cancer chemopreventive activity of terpenoid coumarins from Ferula species. Planta Med, 74: 147-150.
Iranshahi M, Sahbkar A, Takasaki M, Konoshima T, Tokuda H, 2009. Cancer chemopreventive activity of the prenylated coumarin Umbelliprenin in vivo. Eur J Cancer Prev, 18: 412-415.
Iranshahi M, Shahverdi AR, Mirjani R, Amin G, Shafiee A. 2004. Umbelliprenin from Ferula persica roots inhibits the red pigment production in Serratia marcescens. Z Naturforsch, 59: 506-508.
Jabrane A, Ben Jannet H, Mighri Z, Mirjolet JF, Duchamp O, Harzallah-Skhiri F, Lacaille-Dubois MA. 2010. Two new sesquiterpene derivatives from the Tunisian endemic Ferula tunetana Pom. Chem Biodivers, 7: 392-399.
Khaghanzadeh N, Mojtahedi Z, Ramezani M, Erfani N, Ghaderi A. 2012. Umbelliprenin is cytotoxic against QU-DB large cell lung cancer cell line but anti-proliferative against A549 adenocarcinoma cells. Daru. 30:69.
Khorramizadeh MR, Esmail-Nazari Z, Zarei-Ghaane Z, Shakibaieb M, Mollazadeh-Moghaddamd K, et al. 2010. Umbelliprenin-coated Fe3O4 magnetite nanoparticles. Antiproliferation evaluation on human Fibrosarcoma cell line (HT-1080) Fibrosarcoma cell line. Mater Sci Eng C Biomim Mater Sens Syst, 30: 1038-1042.
Krishnan P, Yan K. J, Windler D et al. 2009. Citrus auraptene suppresses cyclin D1 and significantly delays N-methyl nitrosourea induced mammary carcinogenesis in female Sprague-Dawley rats. BMC Cancer, 9: 259.
Li J, Mahdi F, Du L, Jekabsons MB, Zhou YD, Nagle DG. 2013. Semisynthetic studies identify mitochondria poisons from botanical dietary supplements--geranyloxycoumarins from Aegle marmelos. Bioorg Med Chem. 21:1795-803.
Ma CM, Winsor L, Daneshtalab M. 2007. Quantification of spiroether isomers and Herniarin of different parts of Matricaria matricarioides and flowers of Chamaemelum nobile. Phytochem Anal, 18: 42-49.
Mares D, Romagnoli C, Bruni A. 1993. Antidermatophytic activity of Herniarin in preparations of Chamomilla recutita (L) Rauschert Plantes. Med Phytother, 26: 91-100.
Michael O, 2000. The biochemistry of apoptosis. Nature, 407: 770-776.
Mosmann T. 1983. Rapid colorimetric assay for cellular growth and survival. application to proliferation and cytotoxicity assays. J Immunol Methods, 65: 55-63.
Mousavi SH, Tayarani NZ, Parsaee H. 2010. Protective Effect of Saffron Extract and Crocin on Reactive Oxygen Species-Mediated High Glucose-Induced Toxicity in PC12 Cells. Cell Mol Neurobiol, 30: 185-191.
Mousavi SH, Tayarani-Najaran Z, Hersey P. 2008. Apoptosis. from signaling pathways to therapeutic tools. Iran J Basic Med Sci, 11: 121-142.
Murakami A, Matsumoto K, Koshimizu K, Ohigashi H. 2003. Effects of selected food factors with chemopreventive properties on combined lipopolysaccharide- and interferon-g-induced IкB degradation in RAW2647 macrophages. Cancer Lett, 195: 17-25.
Murakami A, Nakamura Y, Ohto Y, Yano M, Koshiba T, et al. 2000. Suppressive effects of citrus fruits on free radical generation and nobiletin. an anti-inflammatory polymethoxyflavonoid. BioFactors, 12: 187-192.
Murakami A, Nakamura Y, Tanaka T, Kawabata K, Takahashi D, et al. 2000. Suppression by citrus Auraptene of phorbol ester- and endotoxin-induced inflammatory responses. role of attenuation of leukocyte activation. Carcinogenesis, 21: 1843-1850.
Murakami A, Wada K, Ueda N et al., 2000. In vitro absorption and metabolism of a citrus chemopreventive agent, auraptene, and its modifying effects on xenobiotic enzyme activities in mouse liver. Nutrition and Cancer, 36:191-199.
Musa MA, Cooperwood JS, Khan MO. 2008. A review of coumarin derivatives in pharmacotherapy of breast cancer. Curr Med Chem, 15: 2664-2679.
Patil J, Kim J, Jayaprakasha G.K. 2010. Berberine induces apoptosis in breast cancer cells (MCF-7) through mitochondrial–dependent pathway. Eur J Pharmacol, 645:70-78
Poncet KM, G Kroemer. 2002. Chemotherapy. targeting the mitochondrial cell death pathway. Oncogene,21: 8786-8803.
Sakata K, Hara A, Hirose Y, Yamada Y, Kuno T, et al. 2004. Dietary supplementation of the Citrus antioxidant Auraptene inhibits NN-Diethylnitrosamine-Induced rat hepatocarcinogenesis. Oncology, 66: 244-252.
Scott EN, Gescher AJ, Steward WP, Brown K, 2009. Development of dietary phytochemical chemopreventive agents biomarkers and choice of dose for early clinical trials. Cancer Prev Res, 6: 525-530.
Shahverdi AR, Saadat F, Khorramizadeh MR, Iranshahi M, Khoshayand MR. 2006. Two matrix metalloproteinases inhibitors from Ferula persica var persica. Phytomedicine, 13: 712-717.
Simstein R, Burow M, Parker A, Weldon C, Beckman B. 2003. Apoptosis, chemoresistance and breast cancer. insights from the MCF-7 cell model system. Exp Biol Med, 9: 995-1003.
Tanaka T, Kawabata K, Kakumoto M, Hara A, Murakami A, et al. 1998. Citrus Auraptene exerts dose-dependent chemopreventive activity in rat large bowel tumorigenesis. the inhibition correlates with suppression of cell proliferation and lipid peroxidation and with induction of phase II drug-metabolizing enzymes. Cancer Res, 58: 2550-2556.
Tanaka T, Kohno H, Murakami M, Kagami S, El-Bayoumy K. 2000. Suppressing effects of dietary supplementation of the organoselenium 14-phenylenebis methylene. Cancer Res, 60: 3713-3716.
Tanaka T, Sugiura H, Inaba R, Nishikawa A, Murakami A, et al. 1999. Immunomodulatory action of citrus Auraptene on macrophage functions and cytokine production of lymphocytes in female BALB/c mice. Carcinogenesis, 20: 1471-1476.
Tayarani-Najaran Z, Mousavi SH, Asili J, Emami SA. 2010. Growth-inhibitory effect of Scutellaria lindbergii in human cancer cell lines. Food Chem Toxicol, 48: 599-604.
Xu W, Cheng M, Lao Y, Wang X, Wu J, Zhou L, Zhang Y, Xu H, Xu N. 2015. DNA damage and ER stress contribute to oblongifolin C-induced cell killing in Bax/Bak-deficient cells. Biochem Biophys Res Commun. 13: 300-306.
Yamada Y, Okamoto M, Kikuzaki H, Nakatani N. 1997. Spasmolytic activity of Auraptene analogs. Biosci Biotechnol, 61: 740-742.