In vitro anti-proliferative activity of clove extract on human gastric carcinoma

Document Type : Original paper


1 Medical Plants Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran.

2 Clinical Biochemistry Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran.

3 Cellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran.


Background and objectives: Cancer cell resistance to common chemotherapy agents is on rise. Plants are considered valuable sources of herbal drugs for cancer therapy. The present study was conducted to investigate the in vitro antioxidant, anti-proliferative, and apoptosis-inducing properties of clove (Syzygium aromaticum L.) extract in human gastric carcinoma (AGS). Methods: Crude ethanol extract of S. aromaticum dried buds was prepared and  in vitro anti-proliferative effects of the extract on AGS and normal Human dermal fibroblasts (HDF) cell lines were studied by MTT assay. To examine apoptosis induction, AGS cells were incubated with IC50 concentrations of the extract, stained with propidium iodide (PI) and annexin V-fluorescein isothiocyanate (FITC), and analyzed by flow cytometry. Antioxidant activity and total phenolics and flavonoids contents were evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, Folin-Ciocalteu method, and aluminum chloride colorimetric method, respectively. Results: The IC50 of DPPH and total phenolics and flavonoids contents of the extract were 10.05±1.93 μg/mL, 225.6±40 mg GAE/g, and 29.30±2.35 mgRUT/g, respectively. The IC50 of the extract against HDFs was 649 µg/mL, higher than AGS cells, which was 118.7 g/mL at 48 h after treatment. Flow cytometric analysis showed that the extract induced cell apoptosis. Conclusions: Crude ethanol S. aromaticum extract had high total phenolics content, and suppressed the proliferation of human gastric cancer cells, likely due to apoptosis induction. Further studies should be conducted to determine the mechanisms of its anticancer effects.


[1] Torre LA, Siegel RL, Ward EM, Jemal A. Global cancer incidence and mortality rates and trends an update. Cancer Epidemiol Biomarkers Prev. 2016; 25(1): 16-27.
[2] Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A. Global cancer statistics, 2012. CA Cancer J Clin. 2015; 65(2): 87-108.
[3] Meyer HJ, Wilke H. Treatment strategies in gastric cancer. Dtsch Arztebl Int. 2011; 108(41): 698-706.
[4] Wagner AD, Unverzagt S, Grothe W, Kleber G, Grothey A, Haerting J, Fleig WE. Chemotherapy for advanced gastric cance. Cochrane Database Syst Rev. 2010; Article ID CD004064.
[5] Van Cutsem E. The treatment of advanced gastric cancer: new findings on the activity of the taxanes. Oncologist. 2004; 9(S2): 9-15.
[6] Asmarian N, Kavousi A, Salehi M, Mahaki B. Mapping of stomach cancer rate in Iran using area-to-area poisson kriging. J Health Syst Res. 2012; 9(3): 681-687.
[7] Ajani JA. Evolving chemotherapy for advanced gastric cancer. Oncologist. 2005; 10(S3): 49-58.
[8] Kaur R, Kapoor K, Kaur H. Plants as a source of anticancer agents. J Nat Prod Plant Resour. 2011; 1(1): 119-124.
[9] Cragg GM, Newman DJ. Plants as a source of anti-cancer agents. J Ethnopharmacol. 2005; 100(1): 72-79.
[10] Moradi MT, Karimi A, Alidadi S. In vitro antiproliferative and apoptosis-inducing activities of crude ethyle alcohole extract of Quercus brantii L. acorn and subsequent fractions. Chin J Nat Med. 2016; 14(3): 196-202.
[11] Sewell RD, Rafieian-Kopaei M. The history and ups and downs of herbal medicines usage. J Herbmed Pharmacol. 2014; 3(1): 1-3.
[12]  Moradi MT, Rafieian-Koupaei M, Imani-Rastabi R, Nasiri J, Shahrani M, Rabiei Z, Alibabaei Z. Antispasmodic effects of yarrow (Achillea millefolium L.) extract in the isolated ileum of rat. Afr J Tradit Complement Altern Med. 2013; 10(6): 499-503.
[13] Karimi A, Mohammadi-Kamalabadi M, Rafieian-Kopaei M, Amjad L. Determination of antioxidant activity, phenolic contents and antiviral potential of methanol extract of Euphorbia spinidens Bornm (Euphorbiaceae). Trop J Pharm Res. 2016; 15(4): 759-764.
[14] Moradi MT, Gatreh-Samani K, Farrokhi E, Rafieian-Koupaei M, Karimi A. The effects of Purslane (Portulaca oleracea L.) on serum level of lipids, lipoproteins and paraoxanase 1(PON1) activity in hypercholesterolemia patients. Life Sci J. 2012; 9(4): 5548-5552.
[15] Mirhoseini M, Moradi MT, Asadi-Samani M. Traditionally used medicinal plants in the treatment of kidney stone: a review on ethnobotanical studies in Iran. Ambient Sci. 2016; 3(2): 16-21.
[16] Moradi MT, Rafieian-Kopaei M, Karimi A. A review study on the effect of Iranian herbal medicines against in vitro replication of herpes simplex virus. Avicenna J Phytomed. 2016; 6(5): 506-515.
[17] Mansouri E, Asadi-Samani M, Kooti W, Ghasemiboroon M, Ashtary-Larky D, Alamiri F, Afrisham R, Hasanzadeh Noohi Z. Anti-fertility effect of hydro-alcoholic extract of fennel (Foeniculum vulgare Mill) seed in male Wistar rats. J Vet Res. 2016; 60(3): 357-363.
[18] Moradi B, Heidari-Soureshjani S, Asadi-Samani M, Yang Q, Saeedi-Boroujeni A. Efficacy and mechanisms of medicinal plants as immunotherapy in treatment of allergic rhinitis: a systematic review. Int J Pharm Sci Res. 2017; 8(5): 1892-1899.
[19] Moradi MT, Karimi A, Rafieian-Kopaei M, Fotouhi F. In vitro antiviral effects of Peganum harmala seed extract and its totalalkaloids against Influenza virus. Microb Pathog. 2017; 110: 42-49.
[20] Karimi A, Moradi MT, Alidadi S, Hashemi L. Anti-adenovirus activity, antioxidant potential, and phenolic content of black tea (Camellia sinensis Kuntze) extract. J Complement Integr Med. 2016; 13(4): 357-363.
[21] Ahmad S, Latif A, Qasmi IA. Effect of 50% ethanolic extract of Syzygium aromaticum (L.) Merr. & Perry.(clove) on sexual behaviour of normal male rats. BMC Complement Altern Med. 2004; Article ID 534794.
[22] Zheng GQ, Kenney PM, Lam LK. Sesquiterpenes from clove (Eugenia caryophyllata) as potential anticarcinogenic agents. J Nat Prod. 1992; 55(7): 999-1003.
[23] Tanaka T, Orii Y, Nonaka G, Nishioka  I. Tannins and related compounds. CXXIII. Chromone, acetophenone and phenylpropanoid glycosides and their galloyl and/or hexahydroxydiphenoyl esters from the leaves of Syzygium aromaticum MERR. et PERRY. Chem Pharmaceut Bull. 1993; 41(7): 1232-1237.
[24] Umehara K, Takagi R, Kuroyanagi M, Ueno A, Taki T, Chen YJ. Studies on differentiation-inducing activities of triterpenes. Chem Pharmaceut Bull. 1992; 40(2): 401-405.
[25] Farag R, Badei A, El Baroty G. Influence of thyme and clove essential oils on cottonseed oil oxidation. J Am Oil Chem Soc. 1989; 66(6): 800-804.
[26] Farag R, Badei A, Hewedi F, El-Baroty G. Antioxidant activity of some spice essential oils on linoleic acid oxidation in aqueous media. J Am Oil Chem Soci. 1989; 66(6): 792-799.
[27] Miyazawa M, Hisama M. Antimutagenic activity of phenylpropanoids from clove (Syzygium aromaticum). J Agric Food Chem. 2003; 51(22): 6413-6422.
[28] Moon JH, Terao J. Antioxidant activity of caffeic acid and dihydrocaffeic acid in lard and human low-density lipoprotein. J Agric Food Chem. 1998; 46(12): 5062-5605.
[29] Folin O, Ciocalteu V. On tyrosine and tryptophane determinations in proteins. J Biol Chem. 1927; 73(2): 627-650.
[30] Karimi A, Moradi MT. Total phenolic compounds and in vitro antioxidant potential of crude methanol extract and the correspond fractions of Quercus brantii L. acorn. J Herbmed Pharmacol. 2015; 4(1): 35-39.
[31] Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983; 65(1-2): 55-63.
[32] Surh YJ. NF-κB and Nrf2 as potential chemopreventive targets of some anti-inflammatory and antioxidative phytonutrients with anti-inflammatory and antioxidative activities. Asia Pac J Clin Nutr. 2008; 17(S1): 269-272.
[33] Williamson G, Manach C. Bioavailability and bioefficacy of polyphenols in humans. II. Review of 93 intervention studies.  Am J Clin Nutr. 2005; 81(S1): 243-255.
[34] Sabu M, Kuttan R. Anti-diabetic activity of medicinal plants and its relationship with their antioxidant property. J Ethnopharmacol. 2002; 81(2): 155-160.
[35] Al-Rashidi W, Mat Supri NN, Manshoor N. Cytotoxic activity of crude extract from Costus malortieanus (Costaceae). Am Eurasian J Toxicol Sci. 2011; 3(2): 63-66.
[36] Treasure T, Fallowfield L, Farewell V, Ferry D, Lees B, Leonard P, Macbeth F, Utley M. Pulmonary metastasectomy in colorectal cancer: time for a trial. Eur J Surg Oncol. 2009; 35(7): 686-689.
[37]  Dwivedi V, Shrivastava R, Hussain S, Ganguly C, Bharadwaj M. Comparative anticancer potential of clove (Syzygium aromaticum)-an Indian spice-against cancer cell lines of various anatomical origin. Asian Pac J Cancer Prev. 2011; 12(8): 1989-1993.
[38] Aisha AF, Abu-Salah KM, Alrokayan SA, Siddiqui MJ, Ismail Z, Majid AMSA. Syzygium aromaticum extracts as good source of betulinic acid and potential anti-breast cancer. Rev Bras Farmacogn. 2012; 22(2): 335-343.
[39] Banerjee S, Panda CK, Das S. Clove (Syzygium aromaticum L.), a potential chemopreventive agent for lung cancer. Carcinogenesis. 2006; 27(8): 1645-1654.
[40] Aggarwal BB, Shishodia S. Molecular targets of dietary agents for prevention and therapy of cancer. Biochem Pharmacol. 2006; 71(10): 1397-1421.
[41] Naik P, Karrim J, Hanahan D. The rise and fall of apoptosis during multistage tumorigenesis: down-modulation contributes to tumor progression from angiogenic progenitors. Genes Dev. 1996; 10(17): 2105-2116.
[42] Rice-Evans C, Miller N, Paganga G. Antioxidant properties of phenolic compounds. Trends Plant Sci. 1997; 2(4): 152-159.
[43] Saleem A, Husheem M, Härkönen P, Pihlaja K. Inhibition of cancer cell growth by crude extract and the phenolics of Terminalia chebula retz. fruit. J Ethnopharmacol. 2002; 81(3): 327-336.
[44] Miliauskas G, Venskutonis P, Van Beek T. Screening of radical scavenging activity of some medicinal and aromatic plant extracts. Food Chem. 2004; 85(2): 231-237.
[45] Kähkönen MP, Hopia AI, Vuorela HJ, Rauha JP, Pihlaja K, Kujala TS, Heinonen M. Antioxidant activity of plant extracts containing phenolic compounds. J Agric Food Chem. 1999; 47(10): 3954-3962.