Hindawi Publishing Corporation Evidence-Based Complementary and Alternative Medicine Volume 2011, Article ID 765029, 11 pages doi:10.1155/2011/765029 Research Article Anticancer Potential of Aqueous Ethanol Seed Extract of Ziziphus mauritiana against Cancer Cell Lines and Ehrlich Ascites Carcinoma Tulika Mishra, 1 Madhu Khullar, 2 and Aruna Bhatia 1 1 Immunology and Immunotechnology Laboratory, Department of Biotechnology, Punjabi University, Patiala, Punjab 147 002, India 2 Experimental Medicine & Biotechnology, Post Graduate Institute of Medical Education & Research, Chandigarh 160 012, India Correspondence should be addressed to Aruna Bhatia, tulika chd@yahoo.com Received 9 May 2010; Revised 9 May 2010; Accepted 10 July 2010 Copyright © 2011 Tulika Mishra et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Ziziphus mauritiana (Lamk.) is a fruit tree that has folkloric implications against many ailments and diseases. In the present study, anticancer potential of seed extract of Ziziphus mauritiana in vitro against different cell lines (HL-60, Molt-4, HeLa, and normal cell line HGF) by MTT assay as well as in vivo against Ehrich ascites carcinoma bearing Swiss albino mice was investigated. The extract was found to markedly inhibit the proliferation of HL-60 cells. Annexin and PI binding of treated HL-60 cells indicated apoptosis induction by extract in a dose-dependent manner. The cell cycle analysis revealed a prominent increase in sub Go population at concentration of 20 μg/ml and above. Agarose gel electrophoresis confirmed DNA fragmentation in HL-60 cells after 3 h incubation with extract. The extract also exhibited potent anticancer potential in vivo. Treatment of Ehrlich ascites carcinoma bearing Swiss albino mice with varied doses (100–800 mg/kg b.wt.) of plant extract significantly reduced tumor volume and viable tumor cell count and improved haemoglobin content, RBC count, mean survival time, tumor inhibition, and percentage life span. The enhanced antioxidant status in extract-treated animals was evident from decline in levels of lipid peroxidation and increased levels of glutathione, catalase, and superoxide dismutase. 1. Introduction The plant-derived compounds have always been an impor- tant source of medicines for various diseases and have received considerable attention in recent years due to their diverse pharmacological properties including cytotoxic and cancer chemopreventive effects [1]. During the last few years, novel chemopreventive agents of natural origin have been targeted with fruits and vegetables being a key interest due to high content of bioactive compounds [2]. Cancer is the second leading cause of death all over the world [3]. According to World Health Organization, more than 10 million new cases of cancer are diagnosed every year, and the statistical trends indicate that this number would double by 2020 [4]. Cancer is the uncontrolled growth and spread of abnormal cells, associated with dysregulation of apoptosis, a programmed cell death. Most of the current anticancer drugs are derived from plant sources, which act through different pathways converging ultimately into activation of apoptosis in cancer cells leading to cell cytotoxicity. Ziziphus mauritiana commonly known as Indian jujube is a fruit tree belonging to family Rhamnaceae. Traditionally, the fruit has been used as anodyne, sedative, tonic anticancer, potent wound healer, applied on cuts and ulcers and has also been used against asthma [5, 6]. The extracts from fruits [7], leaves [8, 9], and seeds [10] of Ziziphus mauritiana have been reported to exhibit antioxidant activity, whereas bark [11, 12] and pulp [13] are reported to possess cytotoxicity against different cancer cell lines. Keeping above in view, the present study was aimed at investigating the effect of aqueous-ethanolic seed extract of Ziziphus mauritiana against different cancer cell lines in vitro and against Ehrlich ascites carcinoma in vivo. 2. Materials and Methods 2.1. Chemicals and Reagents. 3-(4,5-dimethylthiazole-2- yl)- 2,5-diphenyltetrazolium bromide (MTT), RPMI-1640, L- glutamine, penicillin, streptomycin, HEPES, 2-mercaptoeth- anol, propidium iodine (PI), DNase free RNase, proteinase