Cytotoxic effect of Zataria multiflora on cervical cancer cell line (HeLa) and normal fibroblast cells

Maryam JANİTERMİ, Farkhondeh NEMATİ
1.838 356


Abstract. In this study, cytotoxic effects caused by Zataria multifloraessential oil, an Iranian species of family of Laminaceae, in human cervical cancer cell line (HeLa) and human fibroblast cells was investigated. The HeLa cells were seeded in 96-well culture plates in the presence and absence of different concentrations of Z. multifloraessential oil(0, 6.25, 12.5, 25, 50, 100, 200 and 400 µg/ml)for 24, 48 and 72 hours using MTT assay. Results showed thatessential oil of Z. multiflorashowed strong inhibitory effects on HeLa cells when compared to control. The cytotoxicity effect after 72h incubation was greater than 24 and 48h and the highest cytotoxicity of Z. multifloraessential oil was related to 400 μg/ml (100.08%) . Also results showed that more effect of Z. multifloraessenceon the growth inhibitory of normal fibroblast cell was observed at 100 μg/ml after 48h (98.82%).


zataria multiflora, cytotoxic effect, HeLa cells, fibroblast cells, MTT assay

Full Text:



Butler, M.S.(2004),“The role of natural products chemistry in drug discovery,”Journal of Natural Products 67, pp. 2141–2153.

Behtash, N., Ghaemmaghami, F., Ayatollahi, H., et al (2005),“A casecontrol study to evaluate urinary tract complications inradical hysterectomy,”World J Surg Oncol, 3,pp. 12.

Behtash, N., Mousavi, A., Mohit, M., et al (2003),“Simplehysterectomy in the presence of invasive cervical cancer inIran,”Int J Gynecol Cancer, 13, pp.177-81.

Behtash, N., Nazari, Z., Ayatollahi, H., et al (2006), Neoadjuvantchemotherapy and radical surgery compared to radicalsurgery alone in bulky stage 11b-11A cervical cancer,”Eur JSurg Oncol, 32,pp.1226-30.

Mousavi, A. and Karimi-Zarchi, M. (2007),“Isolated cervical metastasisof breast cancer: a case report and literature review,”J LowGenit Tract Dis, 11,pp. 276-8.

Ghaemmaghami, F., Karimi-Zarchi, M., Mousavi, A. et al (2008), “Results of cervical cone excision biopsy in Iran,”Asian Pac J Cancer Prev, 9, pp.45-7.

Mousavi, A., Karimi- Zarchi, M., Gilani, M.M., et al (2008),“Radical hysterectomy in the elderly,”World J Surg Oncol, 6, pp.38.

Talib, H. W.(2011), “Chapter 6: Anticancer and antimicrobial potential of plant-derived natural products. P: 141-158. University of Applied Science, Jordan. Phytochemicals - bioactivities and impact on health,”Edited by Prof. Iraj Rasooli. Pp: 388.

Chiu, H.F. and Wu, Y.C.(2002),“Cytotoxic constituents of the stem bark of Neolitsea acuminatissima,” Journal of Natural Products, 65, pp.255-258.

Cragg, G.M. and Newman, D.J.(1999),“Discovery and development of antineoplastic agents from natural sources,” Cancer Investigation, 17, pp.153-163.

Katsube, N., Iwashita, K., Tsushida, T., Yamaki, K. and Kobori, M.(2003),“Induction of apoptosis in cancer cells by Bilberry (Vaccinium myrtillus) and the anthocyanins,” Journal of Agricultural and Food Chemistry, 51, pp. 68-75.

Mukherjee, A.K., Basu, S., Sarkar, N. and Ghosh, A.C.(2001),“Advances in cancer therapy with plant based natural products,” Current Medicinal Chemistry, 8, pp. 1467-1486.

Richardson, M.A.(2001),“Biopharmacologic and herbal therapies for cancer: research update from NCCAM,” Journal of Nutrition, 131, pp. 3037-3040.

Tatman, D. and Mo, H.(2002),“Volatile isoprenoid constituents of fruits, vegetables and herbs cumulatively suppress the proliferation of murine B16 melanoma and human HL-60 leukemia cells,” Cancer Letters, 175, pp.129-139.

Wargovich, M.J., Woods, C., Hollis, D.M. and Zander, M.E.(2001),“Herbals, cancer prevention and health,” Journal of Nutrition, 131, pp.3034-3036.

Popov, A.M., Atopkina, L.N., Uvarova, N.I. and Elyakov, G.B.(2001),“The antimetastatic and immunomodulating activities of ginseng minor glycosides,” Doklady Biochemistry and Biophysics, 380, pp.309-312.

Ruffa, M.J., Ferraro, G., Wagner, M.L., Calcagno, M.L., Campos, R.H. and Cavallaro, L.(2001),“Cytotoxic effect of Argentine medicinal plant extracts on human hepatocellular carcinoma cell line,” Journal of Ethnopharmacology, 79, pp. 335-339.

Ali, M.S., Saleem, M., Ali, Z. and Ahmad, V.U. (2000), “Chemistry of Zataria multiflora (Lamiaceae),” Phytochem, 55, pp.933–6.

Hosseinzadeh, H., Ramezani, M. and Salmani, G. (2000), “Antinociceptive, anti- inflammatory and acute toxicity effects of Zataria multiflora Boiss. extracts in mice and rats,” J Ethnopharmacol, 73, pp. 379–85.

Jafarian-Dehkordi, A., Emami, S.A., Saeidi, M. and Sadeghi, H. (2004), “Cytotoxicologic studies of the extracts of iranian Juniperus sabina and Platycladus orientalis on cancer cells,” J Res Med Sci, 5, pp.205–9.

Saeedi-Saravi, S.S. and Shokrzadeh, M. (2008), “The study of hepatic and renal disorders in mice which were administered ethyl acetate extract of plant Sambucus ebulus Intraperituenally (IP) and effect of vitamins C and E on prevention of its disorders,” Toxicol Lett,180, pp.57–58.

Shaffiee, A. and Javidnia, K. (1997), “Composition of essential oil of Zataria multiflora,” Planta Med, 63, pp. 371–2.

Saei-Dehkordi, S.S., Tajik, H., Moradi, M. and Khalighi-Sigaroodi, F. (2010),“Chemical composition of essential oils in Zataria multiflora Boiss. From different parts of Iran and their radical scavenging and antimicrobial activity,” Food Chemistry and Toxicology 48, pp.1562-1567.

Aeschbach, R., Loliger, J., Scott, B.C., Murcia, A., Butler, J., Halliwell, B. and Aruoma, O.I.(1994),“Antioxidant actions of thymol, carvacrol, 6-gingerol, zingerone and hydroxytyrosol,” Food and Chemical Toxicology, 32, pp. 31–36.

Mastelic, J., Jerkovic, I., Blazevic, I., Poljak-Blazi, M., Borovic, S., Ivancic-Bace, I., Smrecki, V., Zarkovic, N., Brcic-Kostic, K., Vikic-Topic, D. and Muller, N.(2008), “Comparative study on the antioxidant and biological activities of carvacrol, thymol, and eugenol derivatives,” Journal of Agricultural and Food Chemistry, 56, pp. 3989–3996.

Kallistratos, G., Evangelou, A., Agnantis, N., Fasske, E., Karkabounas, S. and Donos, A.(1994), “Enhancement of the antineoplastic effect of anticarcinogens on benzo[a]pyrene- treated Wistar rats, in relation to their number and biological activity,” Cancer Letters, 82, pp. 153–165.

Evangelou, A., Kalpousos, G., Karkabounas, S., Liasko, R., Nonni, A., Stefanou, D. and Kallistratos, G.(1997), “Dose-related preventive and therapeutic effects of antioxidants - anticarcinogens on experimentally induced malignant tumors in Wistar rats,” Cancer Letters, 115, pp.105–111.

Liasko, R., Kabanos, T., Karkabounas, S., Malamas, M., Tasiopoulos, A., Stefanou, D., Collery, P. and Evangelou, A.(1998),“Beneficial effects of a Vanadium complex with cysteine administrated at low doses of benzo[a]pyrene - induced leiomyosarcomas in Wistar rats,” Anticancer Research, 18, pp. 3609–3613.

Karkabounas, S., Binolis, J., Zelovitis, J., Kotsis, N., Charalabopoulos, A., Avdikos, A., Zouridakis, A., Liasko, R., Giannakopoulos, X. and Charalabopoulos, K.(2002),“Inhibition and modification of benzo[a]pyrene-induced chemical carcinogenesis by ascorbic acid alone or in combination with a-tocopherol in Wistar rats,” Experimental Oncology, 24, pp. 274–278.

He, L., Mo, H., Hadisusilo, S., Qureshi, A.A. and Elson, C.E. (1997), “Isoprenoids suppress the growth of murine B16 melanomas in vitro and in vivo,” J Nutr 127, pp. 668– 674.

Horvathova, E., Turcaniova, V. and Slamenova, D.(2007),“Comparative study of DNA damaging and DNA-protective effects of selected components of essential plant oils in human leukemic cells K562,” Neoplasma, 54, pp.478–483.

Karkabounas, S., Kostoula, O.K., Daskalou, T., Veltsistas, P., Karamouzis, M., Zelovitis, I., Metsios, A., Lekkas, P., Evangelou, A.M., Kotsis, N. and Skoufos, I.(2006),“ Anticarcinogenic and antiplatelet effects of carvacrol,” Experimental Oncology, 28, pp.121–125.

Koparal, A.T. and Zeytinoglu, M.(2003), Effects of carvacrol on a human non-small cell lung cancer (NSCLC) cell line A549,” Cytotechnology, 43, pp. 149–154.

Lampronti, I., Saab, A.M. and Gambari, R.(2006), “Antiproliferative activity of essential oils derived from plants belonging to the Magnoliophyta division,” International Journal of Oncology, 29, pp. 989–995.

Arunasree, K.M.(2010), “Anti-proliferative effects of carvacrol on a human metastatic breast cancer cell line, MDA-MB 231,” Phytomedicine, 17, pp. 581–588.

Phelan, M.C. (1998), “Basic Techniques for Mammalian to Cell Tissue Culture,” Current protocols in cell biology; 7, pp. 1-10.

Green, J.E. (2003), “Mouse models of human breast cancer: evolution or convolution,” Breast Cancer Res. 5(1), pp.1.

Arya, V., Kashyap, C.P.,Bandana, T.,Shiksha, S.,Sweta, K.,Verma,P. and Sharma, S. (2011),“Human cancer cell lines A brief communication, J. Chem. Pharm. Res. 3(6), pp.514-520

Spavieri, J., Allmendinger, A., Kaiser, M., Casey, R.,Hingley-Wilson S.and Lalvani, A. (2010),“Antimycobacterial, antiprotozoan and cytotoxic potential of twenty-one brown algae (Phaeophyceae) from British and Irish waters,” Phytother. Res. 24 : 1724-29.

Alizadeh, A. and Shaabani, M. (2014), “Essential oil composition, total phenolic content and antioxidant activities of Iranian Zataria multiflora Boiss,” International Journal of Biosciences. 4 (4), pp. 97-104.

Ebrahimzadeh, H., Yamini, Y., Sefidkon, F., Chaloosi, M. and Pourmortazavi, S.M. (2003), “Chemical composition of the essential oil and supercritical CO2 extracts of Zataria multiflora Boiss,” Food Chem., 83, pp. 357-361.

Sharififar, F., Moshafi, M.H., Mansouri, S.H., Khodashenas, M. and Khoshnoodi, M. (2007), “In vitro evaluation of antibacterial and antioxidant activities of the essential oil and methanol extract of endemic Zataria multiflora Boiss,” Food Control, 18, pp. 800-805.

Alipour-eskandani, M. (2011), “Effect of Zataria multifloraBoiss essential oil on the growth of bacillus cereus in a commercial chicken soup,” Book of proceedings 6, pp. 389- 392.

Stammati, A., Bonsi, P., Zucco, F., Moezelaar, R., Alakomi, H.L. and Von-Wright, A. (1999), “Toxicity of selected plant volatiles in microbial and mammalian short-term assays,” Food Chem Toxicol 37, pp. 813–823.

Zeytinoglu, M., Aydin, S., Ztrk, Y., Hsn, K. and Baser, C. (1998), “Inhibitory effects of carvacrol on DMBA induced pulmonary tumorigenesis in rats,” Acta Pharmaceutica Turcica 40, pp. 93–98.

Zeytinoglu, H., Incesu, Z. and Baser, K. (2003), “Inhibition of DNA synthesis by Carvacrol in mouse myoblast cells bearing a human N-RAS oncogene,” Phytomedicine 10, pp. 292– 299.

Sylvester, P.W. (2011), “Optimization of the tetrazolium dye (MTT) colorimetric assay for cellular growth and viability,” Methods Mol Biol, 716, pp.157-68.

Maioli, E., Torricelli, C., Fortino, V., Carlucci, F., Tommassini, V. and Pacini, A. (2009), “Critical appraisal of the MTT assay in the presence of rottlerin and uncouplers,” Biol Proced Online, 3(11), pp. 227-40.

Zamanian-Azodi, M., Rezaie-Tavirani, M., Heydari- Kashal, S., kalantari, S., Dalilan, S. and Zali, H. (2012), “Proteomics analysis of MKN45 cell line before and after treatment with Lavender aqueous extract,” Gastroenterol Hepatol Bed Bench, 5, pp.35-42.