American Journal of Cancer Prevention, 2015, Vol. 3, No. 2, 45-50 Available online at http://pubs.sciepub.com/ajcp/3/2/4 © Science and Education Publishing DOI:10.12691/ajcp-3-2-4 Anti-proliferative, Cytotoxicity and Anti-oxidant Activity of Juglans regia Extract Tajamul Islam Shah 1,* , Ekta Sharma 2 , Gowhar Ahmad Shah 3 1 Department of Research, Jawaharlal Nehru Cancer Hospital and Research Centre, Bhopal, India 2 Department of Botany, Benazeer College of science, Bhopal, India 3 Department of Zoology, Benazeer College of Science and Commerce Bhopal, India *Corresponding author: taju.zoology@gmail.com Received April 01, 2014; Revised April 09, 2014; Accepted April 20, 2015 Abstract Objective: Juglans regia (walnuts), the royal species from Junglandaceae family, well-known for its valuable medicinal uses, their regular consumption may have beneficial effects against oxidative stress mediated diseases including cancer. The present study was aimed to explore the total phenolic content, anti-proliferative and anti-oxidant activity of Juglans regia leaves. Methods: The leaf powder was extracted using different solvents and subjected for phytochemical investigation. The total phenolic contents were determined by the Folin–Ciocalteu method. The extracts comprising a good amount of secondary metabolites especially polyphenols were used to evaluate their antioxidant activity using Fenton’s reagent reaction and DPPH scavenging assay, while cytotoxic and anti-proliferative activity against B16F10 mice melanoma and A375 human melanoma cell were screened using MTT Assay. Observation: Methanolic extract presented the highest total phenolic content (94.39 ± 5.63 mg of GAE/g of extract) as compared to aqueous extract (27.92 ± 1.40 mg of GAE/g of extract). Similarly, methanolic extract presented the highest antioxidant activity (EC 50 of 0.250mg/ml) followed by water extract (EC 50 of 0.325mg/ml) in Fenton’s reaction and 0.199 ± 0.023 and 2.991 ± 0.740, respectively in DPPH assay. The extracts showed concentration dependent growth inhibition activity (IC 50 0.234 and 0.304mg/ml) against B16F10 mice melanoma and A375 human melanoma cell line (IC 50 0.298 and 0.350mg/ml) respectively. The extracts proved least toxic when treated with normal lymphocytes. The results indicate that walnut leaves are an excellent source of antioxidant and anti-cancerous agents and may prove fruitful herbal remedy in near future. However, the extracts proved effective against mice melanoma and human melanoma cells. Despite, more study is required before coming to any conclusion. Keywords: J. regia, anti-oxidant, anti-proliferative, B16F10, A375, Lymphocytes, MTT Cite This Article: Tajamul Islam Shah, Ekta Sharma, and Gowhar Ahmad Shah, “Anti-proliferative, Cytotoxicity and Anti-oxidant Activity of Juglans regia Extract.” American Journal of Cancer Prevention, vol. 3, no. 2 (2015): 45-50. doi: 10.12691/ajcp-3-2-4. 1. Introduction Epidemiological studies have consistently shown that there is a clear significant positive association between regular consumption of fruits, nuts and vegetables, and a reduced incidence of ischemic heart disease and some types of cancer, particularly stomach, oesophagus, lung, oral cavity and pharynx, endometrial, pancreas and colon cancers [1,2,3,4,5]. These antioxidant and chemopreventive properties have been attributed to their high content of natural antioxidants, especially ascorbic acid (vitamin C), tocopherols (vitamin E), b-carotene (provitamin A), anthocyanins and other polyphenols [6,7,8,9]. Several studies showed that phenolic compounds are the major bioactive phytochemicals with human health benefits [10,11,12]. In fact, many authors have reported a direct relationship between total phenolic content and antioxidant activity in numerous seeds, fruits and vegetables [9,10,11,12,13]. The Juglans genus (family Juglandaceae) comprises several species and is widely distributed throughout the world. The walnut tree (Juglans regia L.) is its well-known member, constituting an important species of deciduous trees found primarily in temperate areas and cultivated commercially throughout southern Europe, northern Africa, eastern Asia, United States and western South America. In Portugal, walnut trees can be found all over the country, being the nuts very popular and largely consumed as part of the Mediterranean diet. Nevertheless, not only dry seeds (nuts) are used but also green walnuts, shells, bark, green husks (epicarps) and leaves, which have been used in the cosmetic and pharmaceutical industries [14]. Walnut leaves are considered a source of healthcare com- pounds, and have been widely used in traditional medicine for treatment of skin inflammations, hyperhidrosis and ulcers and for its antidiarrheic, anti- helmintic, antiseptic and astringent prop- erties [15]. In Portugal, as in some other European countries, especially in rural areas, dry walnut leaves are fre- quently used to