Study of cytotoxic effect of Essential oil of Tymus vulgaris L by MTT and Trypan blue assay on breast cancer cell line MCF-7

Subject Areas : Developmental biology of plants and animals , development and differentiation in microorganisms

Sasan Hajebi ¹ , Abdolkarim Chehregani Rad ² , Hossein Lariyazdi ³

1 - Young Researchers and Elite Club, Borujerd Branch, Islamic Azad University, Borujerd, Iran
2 - Department of Biology, Faculty of Science, Bu-Ali Sina University, Hamedan, Iran
3 - Department of Biology, Faculty of Science, Islamic Azad University, Borujerd, Iran

Received: 2018-01-08 Accepted : 2018-04-07 Published : 2017-11-22

Keywords: breast cancer, MTT, Thymus vulgaris L,

Abstract :

Breast Cancer is the most common cancer in the Iranian women. Essential oil of Thymus is containing anticancer compounds including thymol. The aim if this research was studding the effect of thymus essential oil on the breast cancer cell line MCF-7 using MTT and Trypan blue methods, and comparing of its effect with taxol as a well-known anticancer agent. In this research, MCF-7 and Hu-02 cell lines were porched from IBRC institute. The cells were treated by different concentrations of Thymus essential oil and taxol in the times of 24, 48 and 72 hours. The cytotoxicity of the treatments was measured by means of MTT and trypan blue staining. Results showed that Thymus essential oil had cytotoxic effect, in different doses, on the MCF-7 cell line. The highest cytotoxic effect was measured in the group treated by 125 μg/ml of Thymus essential oil. The Thymus essential did not show any cytotoxic effect on the cells of HU-02. Results were also showed that the essential oil can increase the cytotoxicity effect of Taxol on the cancer cell line, thus it seems that is effective in breast cancer therapy; probably, it causes to cell death by apoptosis induction.

References:

[1]. Amirghofran Z., and Karimi, M.H. 2002. Cytotoxic activity of Thymus vulgaris, Achillea Millefolium and Thuja orientalis on different growing cell lines. J Medical Journal of the Islamic Republic of Iran, 15: 149-154.

[2]. Ayesh, B.M., Abed, A.A., and Faris, D.M. 2014. In vitro inhibition of human leukemia THP-1 cells by Origanum syriacum L. and Thymus vulgaris L. extracts. J BMC Research Notes. 7:612620.

[3]. Craig, W.J. 1999. Health promoting properties of common herbs. American J of clinicalnutrition. 70(3): 491-499.

[4]. Dalilan, S., Rezaei-Tavirani, M., Nabiuni, M., Heidari-Keshel, S, Zamanian-Azodi, M., and Zali, H. 2013. Aqueous extract of Lavender angustifolia inhibits lymphocytes proliferation of hodgkin's lymp homaPatients. Iran J Cancer Prev. 4: 201208.

[5]. De Brabandert, M., Geuens, G., Nuydens, R., Willebrords, R., and De Mey, J.1981. Taxol induces the assembly of free microtubules in living cells and blocks the organizing capacity of the centrosomes and kinetochores. Proc Natl Acad Sci USA. 78:5608–5612.

  [6]. Esmaeili-Mahani, S., Falahi, F., and Yaghoobi M.M. 2014. Proapoptotic and Antiproliferative Effects of Thymus caramanicus on Human Breast Cancer Cell Line (MCF-7) and Its Interaction with Anticancer Drug Vincristine. J EvidenceBased Complementary and Alternative Medicine (On line published). http://dx.doi.org/10.1155/2014/893247

[7]. Evandri, M.G., Battinelli, L., Daniele, C., Mastrangelo, S., Bolle, P., Mazzanti, G. 2005. The antimutagenic activity of Lavandula angustifolia (lavender) essential oil in the bacterial reverse mutation assay. 2005. J Food and Chemical Toxicology 43: 1381 -1387.

[8]. Guo, J., Bourre, L., Soden, D.M., O’Sullivan, G.C., O’Driscoll C. 2011. Non-Viral Technologies Knockdown the Barriers to siRNA Delivery and Achieve the Next Generation of Cancer Therapeutics, J Biotechnol Adv. 29(4):40217.

[9]. Harirchi, I., Karbakhsh, M., Kashefi, A., Momtahen, A.J. 2004. Breast Cancer in Iran: Results of a Multi-Center Study. Asian Pac J Cancer Prev. 5(1):24-7.

[10]. Harirchi, I., Ebrahimi, M., Zamani, N., Jarvandi, S., Montazeri, A. 2000. Breast Cancer in Iran: a Review of 903 Case Records. J Public Health. 114(2):143-5.

[11]. Hoffman, E. Cancer and the search for selective biochemical inhibitors. 1999. London: Boca Raton: 95-97.

[12]. Hornick, J.E., Bader, J.R., Tribble, E.K., Trimble, K., Breunig, J.S., Halpin, E.S., Vaughan, K.T., and Hinchcliffe, E.H. 2008. Live-cell analysis of mitotic spindle formation in Taxol-treated cells. Cell Motil Cytoskeleton 65:595–613.

[13]. Joekim, H., Chung, J.H., Lee S.H., Jung, Y.S., Moon, C.H., and Baik, E.J. 2008. Involvement of endogenous prostaglandin- f2 alpha on kainic acid– induced seizureactivity through fp receptor: the mechanism of proconvulsant effects of COX-2 inhibitors. Brain Res. 193: 153-61.

[14]. Katzung, B.G., Masters, S.B., and Terevor, A.J. 1997 Basic and clinical pharmacology. New York, Norwalk.

[15]. Kazemi, M. 2015. Phytochemical Composition of Thymus vulgaris L. Essential Oil. J of Essential Oil Bearing Plants. 18(3): 751-753.

[16]. Kidwia, M., Venkataramanan, R., Mohan, R., Sapra, P. 2002. Cancer chemotherapy and heterocyclic compounds. J Curr Med Chem. 9 (12): 1209-1228.

[17]. Liang, Y.C., Tsai, S.H., Tsai, D.C., and Lin-shiau, S.Y. 2001. Suppression of inducible cyclooxygenase and nitric oxide synthase through activation of peroxisome proliferator– activated receptor –gamma by flavonoids in mouse macrophages . FEBS Lett. 496(1): 12- 18.

[18]. Lixandru, B.E., Dracea, N.O., Dragomirescu, C.C., Dragulescu, E.C., Coldea, I.L., Anton, L., et al. 2010. Antimicrobial activity of plant essential oils against bacterial and fungal species involved in food poisoning and/or food decay. J Roum Arch Microbiol Immunol. 69: 224-30. 

[19]. Lo, A.H., Liang, Y.C., Lin-Shiau, S.Y., Ho, C.T., et al. 2002. Carnosol, an antioxidant in rosemary, suppresses inducible nitric oxide synthase through downregulating nuclear factor-kappaB in mouse macrophages. Carcinogenesis. 23(6): 983-991.

[20]. Mans, D.R.A., and Schwartsmann, G. 2000. Anticancer drug discovery and development in Brazil: targeted plant collection as a rational strategy to acquire candidate anticancer compound. Oncologist. 5: 185-98.

[21]. Mastelic, J., Jerkovic, I., Blazevic, I., Poljak-Blazi, M., Borovic, S., IvancicBace, I., et al. 2008. Comparative study on the antioxidant and biological activities of carvacrol, thymol, and eugenol derivatives. J Agric Food Chem. 56: 3989-96.

[22]. McKay, D.L., and Blumberg, J.B. 2006. Review of the bioactivity and potential health benefits of peppermint tea (Mentha piperita L.). J Phytother. Res. 26: 619- 633.

[23]. Moller, P., Wallin, H., and Knudsen, L.E. 1996. Oxidative stress associated psychological stress and lifestyle factor. Chem Bio Interact. 102: 1-36.

[24]. Mosmann, T. 1998. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J. Immunol. Methods. 65, 55–63.

[25]. Osakabe, N., Yasuda, A., Natsume, M., Sanbongi, C., Kato, Y., Osawa, T., et al., 2002. Rosmarinic acid, a major polyphenolic component of Perilla frutescens, reduces lipopolysaccharide (LPS) induced liver injury in dgalactosamine (d- GalN)- sensitized mice Free Radic. J Biol. Med. 33: 798-806.

[26]. Osakabe, N., Yasuda, A., Natsume, M., and Yoshikawa, T. 2004. Rosmarinic acid inhibits epidermal inflammatory responses: anticarcinogenic effect of Perilla frutescens extract in the murine two-stage skin model. Carcinogenesis. 25: 549-557.

[27]. Talib W H, and Mahasneh A M. 2010. Antiproliferative Activity of Plant Extracts Used Against Cancer in Traditional Medicine. Sci Pharm, 78: 33-45.

[28]. Patricia, P., and Trevor J.M. 1990. Use of the Tetrazolium Assay in Measuring the Response of Human Tumor Cells to Ionizing Radiation, J cancer research. 50: 1392-1396.

[29]. Rezaianzadeh, A., Peacock, J., Reidpath, D., Talei, A., Hosseini, S.V., and Mehrabani, D. 2009. Survival Analysis of 1148 Women Diagnosed with Breast Cancer in Southern Iran. BMC Cancer. (On line published).

[30]. Sadjadi, A., Nouraie, M., Mohagheghi, M.A., Mousavi-Jarrahi, A., Malekezadeh, R., and Parkin, D.M. 2002. Cancer Occurrence in Iran in 2002, an International Perspective. Asian Pac J Cancer Prev. 6(3):359-363.

[31]. Sadiq, B.M., Naz, A., Tauseef, S.M., and Qayyum, M.M.N. 2013. Anti-oncogenic perspectives of spices/herbs. EXCLI J. 12: 1043-1065.

[32]. Singh B, and Lucci A. 2002. Role of cycloxygenase-2 in breast cancer. Surg Res. 108: 173-179.

[33]. Slater, T.F., Sawyer, B., and Strauli U.D. 1963. Studies on cuccinate-Tetrazolinum reductase system III. Point of coupling of four different tetrazolum salt. J Biochim. Biophys. Acta. 77: 383-393.

[34]. Srivastava, V., Negi, A.S., Kuma, J.K.R, Gupta, M.M., and Khanuja, S.P.S, 2005. Plant based anticancer molecules: A chemical and biological profile of some important leads. J Bioorganic & Medicinal Chemistry.13: 582-590.

[35]. Sylvestre, M., Pichette, A., Longtin, A., Nagau, F., and Legault, J. 2006. Essential oil analysis and anticancer activity of leaf essential oil of Croton flavens L. from guadeloupe. J. Ethnopharmacol. 103: 99102.

[36]. Sylvestre, M., Legault, J., Dufour, D., and Pichette, A. 2005. Chemical composition and anticancer activity of leaf essential oil of Myrica gale L. J. Phytomedicine. 12: 299-304.

[37]. Undeger, U., Başaran, A., Degen, G.H., and Başaran, N. 2009. Antioxidant activities of major thyme ingredients and lack of (oxidative) DNA damage in V79 Chinese hamster lung fibroblast cells at low levels of Carvacrol and Thymol. J Food Chem Toxicol. 47: 2037-2043.

[38]. Vigo, E., Cepeda, A., Gualillo, O., and Perez-Fernandez, R. 2004. In-vitro antiinflammatory effect of Eucalyptus globulus and Thymus vulgaris: nitric oxide inhibition in J774A.1 murine macrophages. J Pharm Pharmacol. 56: 257-263.

[39]. Wei, W., Nan, L.i., Meng, L., Yuangang, Z.u. and Efferth, T. 2010. Antibacterial activity and Anticancer activity of Rosmarinus officinalis L. essential oil compared to that of its main components. Molecules. 17: 2704-2713.