• Ahmed R. Abu-raghif Al-Nahrain University/College of Medicine/ Pharmacology Department, Baghdad-Iraq
  • Ghaith A. Jasim Al-Mustansiriyah University/College of Pharmacy/Department of Pharmacology & Toxicology, Baghdad-Iraq
  • Muneer M. Hanoon Thi Qar Health Office/ Thi Qar Poison Control Center



Anti-proliferative activity, Zizyphus spina christi, Methanol extract, RD cell line, MTT assay, DPPH assay


Objective: The aim of this study was to investigate the capability of Zizyphus spina christi methanol extract to inhibit cancer cell line proliferation.

Methods: The leaves of Zizyphus spina christi were extracted by cold maceration method. The anti-proliferative activity of the methanol extract against rhabdomyosarcoma (RD) cell line was tested by 3-(4, 5 Dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The phytochemical constituents were identified by gas chromatography-mass spectrometry (GC-MS) analysis. The antioxidant activity was assessed by measuring free radical scavenging activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay.

Results: The percentage extraction yield for leaves with methanol was 20.64%. The methanol extract showed dose dependent inhibition of RD cell line, the IC50 was 154.44 µg/ml. GC-MS showed the presence of flavonoid fraction and other compounds with antioxidant activity. The methanol extract demonstrated DPPH scavenging activity with IC50 of 33.91 mg/ml.

Conclusion: Methanol extract showed potential anti-proliferative activity against RD cell line, which could be due to its antioxidant activity.


Download data is not yet available.


Lopez-Gomez M, Malmierca E, de Gorgolas M, Casado E. Cancer in developing countries: the next most preventable pandemic. The global problem of cancer. Crit Rev Oncol Hematol 2013;88:117-22.

American Cancer Society. Cancer Facts and fig. 2012. Atlanta: American Cancer Society.

Saab R, Spunt SL, Skapek SX. Myogenesis and rhabdomyosarcoma the Jekyll and Hyde of skeletal muscle. Curr Top Dev Biol 2011;94:197–234.

Ognjanovic S, Linabery AM, Charbonneau B, Ross JA. Trends in childhood rhabdomyosarcoma incidence and survival in the United States, 1975–2005. Cancer 2009;115(18):4218–26.

Cancer Research UK. Side Effects of Cancer Drugs. Available online: http://cancerhelp. cancerresearchuk. Org/about-cancer/treatment/cancer-drugs/side-effects (accessed on 14 February 2013).

Raguz, S, YagueE. Resistance to chemotherapy: New treatments and novel insights into an old problem. Br. J. Cancer 2008;99:387–91.

Yossef HE, Khedr AA, Mahran MZ. Hepatoprotective activity and antioxidant effects of El Nabka (Zizyphus spina christi) fruits on rat hepatotoxicity induced by carbon tetrachloride. Nat. Sci 2011;9:17.

Duke JA. Handbook of Medical Herbs. CRC Press: Boca Raton; 1985.

Asgarpanah J, Haghigat E. Phytochemistry and pharmacologic properties of Ziziphus spina christi (L.) Willd. Afr J Pharm Pharmacol 2012;6:2332-9.

Pathmanathan MK, Uthayarasa K, Jeyadevan JP, Jeyaseelan EC. In vitro antibacterial activity and phytochemical analysis of some selected medicinal plants. Int J Pharm Biol Arch 2010;1:291-9.

Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983;65:55-63.

Mujeeb F, Bajpai P, Pathak N. Phytochemical evaluation, antimicrobial activity and determination of bioactive components from leaves of aegle marmelos. BioMed Res Int 2014;1-11.

Oktay M, Gulcin I, Kufrevioglu OI. Determination of in vitro anti-oxidant activity of funnel (Foeniculum vulgare) seed extracts. Lebensm-Wiss U-Technoli 2003;36:263-71.

Shams KA, Abdel-Azim NS, Saleh IA. Green technology: economically and environmentally innovative methods for extraction of medicinal andaromatic plants (MAP) in Egypt. J Chem Pharm Res 2015;7:1050-74.

Sasidharan S, Chen Y, Saravanan D, Sundram KM, Yoga Latha L. Extraction, isolation and characterization of bioactive compounds from plants’ Extracts. Afr J Traditional Complementary Alternative Med 2011;8:1-10.

Goldin A, Venditti JM, MacDonald JS. Current results of the screening program at the division of cancer treatment, National Cancer Institute. Eur J Cancer 1981;17:129-42.

Venditti JM. Preclinical drug development: rationale and methods. Semin Oncol 1981;8:349-61.

Schmahl D, Berger MR. Possibilities and limitations of antineoplastic chemotherapy: experimental and clinical aspects. Int J Exp Clin Chemother 1988;1:1-11.

Scudiero DA, Shoemaker RH, Paull KD. Evaluation of a soluble tetrazolium/formazan assay for cell growth and drug sensitivity in culture using human and other tumor cell lines. Cancer Res 1988;48:4827-33.

Ahmed R Abu-Raghif, Hayder B Sahib, Muneer M Hanoon. Anti-angiogenic activity of Zizyphus spina christi Leaves Extracts. Int J Pharm Sci Rev Res 2015;35:169-74.

Vijayarathna S, Sasidharan S. Cytotoxicity of methanol extracts of Elaeis guineensis on MCF-7 and Vero cell lines. Asian Pac J Trop Biomed 2012;2:826-9.

Batra P, Sharma AK. Anti-cancer potential of flavonoids: recent trends and future perspectives. Biotechnology 2013;3:439–59.

Ren W, Qiao Z, Wang H, Zhu L, Zhang L. Flavonoids: promising anticancer agents. Med Res Rev 2003;23:519-34.

Harada H, Yamashita U, Kurihara H. Antitumor activity of palmitic acid found as a selective cytotoxic substance in a marine red alga. Anticancer Res 2002;22:2587−90.

Sermakkani M, Thangapandian V. gc-ms analysis of Сassia italica leaf methanol extract. Asian J Pharm Clin Res 2012;5:90-4.

Gopalakrishnan K, Udayakumar R. GC-MS analysis of phytocompounds of Leaf and Stem of Marsilea quadrifolia (L.). Int J Biochem Res Rev 2014;4:517-26.

Aparna V, Dileep KV, Mandal PK. AntiInflammatory property of n-hexadecanoic acid: structural evidence and kinetic assessment. Chem Biol Drug Des 2012;80:434–9.

Jeong WC, Kim KJ, Ju H. Cytoplasmic phospholipase A2 metabolites play a critical role in pulmonary tumor metastasis in mice. Anticancer Res 2010;30:3421-7.

Linkous AG, Yazlovitskaya EM, Hallahan DE. Cytosolic phospholipase A2 and lysophospholipids in tumor angiogenesis. J Natr Cancer Inst 2010;102:1398–412.

Chidambaram V, Niraimathi L, Sudha V. Spectrophotometric, HPTLC and GC-MS studies on selected spice extracts. Int J Pharm Pharm Sci 2015;7:184-90.

Kedare SB, Singh RP. Genesis and development of DPPH method of antioxidant assay. J Food Sci Technol 2011;48: 412–22.

Pryor WA, Houk KN, Foote CS. Free radical biology and medicine: it‟s a gas, man! Am J Physiol: Regul Integr Comp Physiol 2006;291:491–511.

Singh S, Singh RP. In vitro methods of assay of antioxidants: an overview. Food Rev Int 2008;24:392–415.

Al-Jassabi S, Abdullah MS. Extraction, purification and characterization of antioxidant fractions from Zizyphus spina-christi fruits. Am-Eur J Toxicol Sci 2013;5:66-71.

Boeing JS, Barizaol EO, Silva CS, Montanher PF, Almeida VC, Visentainer JV. Evaluation of solvent effect on the extraction of phenolic compounds and antioxidant capacities from the berries: application of principal component analysis. Chem Central J 2014;8:48.

Khaleel. Evaluation of total phenolic content and antioxidant activity of three leaf extracts of Ziziphus spina-christi (Sedr) Grown in Jordan. Br J Med Med Res 2016;14:1-8.

Brum TF, Zadra M, Piana M. HPLC analysis of phenolics compounds and antioxidant capacity of leaves of Vitex megapotamica (Sprengel) Moldenke. Molecules 2013;18: 8342-57.

Halliwell B, Gutteridge JMC. Free radicals in biology and medicine. 3rd edition. Clarendon Press: Oxford, UK; 1999.

Mishra A, Kumar S, Pandey AK. Scientific validation of the medicinal efficacy of Tinospora cordifolia. Sci World J 2013.

Aruoma OI. Nutrition and health aspects of free radicals and antioxidants. Food Chem Toxicol 1994;32:671–83.

Fang YZ, Yang S, Wu G. Free radicals, antioxidants, and nutrition. Nutrition 2002;18:872–9.



How to Cite

Abu-raghif, A. R., G. A. Jasim, and M. M. Hanoon. “ANTI-PROLIFERATIVE ACTIVITY OF ZIZYPHUS SPINA CHRISTI LEAVES METHANOL EXTRACT AGAINST RHABDOMYOSARCOMA (RD) CELL LINE”. International Journal of Pharmacy and Pharmaceutical Sciences, vol. 9, no. 2, Feb. 2017, pp. 279-82, doi:10.22159/ijpps.2017v9i2.15013.



Original Article(s)