A difference of P 0. 05 was con sidered to be statistically significant and the

Dillenia suffruticosa Martelli, which belongs to family Dilleniaceae, is a plant native in Peninsular Malaysia, Kalimantan, Sumatra and Singapore. The evergreen shrub can be found in secondary forest and swampy ground. The supplier KU-0063794 fruit of the plant has the ability to treat cancerous growth. Other traditional use of the plant is to relieve rheumatism. Methanolic extract of D. suffruticosa showed a broad spectrum of antimicrobial activity against Bacillus cereus, Bacillius subtilis, Candida albicans, and Pseudomonas aeruginosa. Water extract of D. suffruticosa also exhibited inhibitory action against replication of dengue virus type 2. Armania et al. reported that extract of D. suffruticosa showed high antioxidant and cytotoxic activities towards various cell lines including Hela, MCF 7, MDA MB 231, A549 and HT 29 cell lines.

In this study, root extract was selected for elaborated study. As the previous study demonstrated that root extract of the plant exhibited the most potent cytotoxic activity, in comparison to fruit, leaf, and flower parts of the plant. The aim of this study was to investigate supplier Lenalidomide the anticancer effect of ethyl acetate of D. suffruticosa in breast cancer cells, MCF 7, and to explore the apoptotic signaling pathway underlying it. Methods Chemicals and reagents Hexane, dichloromethane, ethyl acetate and dimethyl sulfoxide were purchased from FS Chemicals. RPMI 1640 was purchased from Nacalai Tesque. Fetal bovine serum, trypsin, streptomycin and penicillin were obtained from PAA Laboratories GmBH. 3 2,5 diphenyltetra zolium bromide, propidium iodide and RNAse A were purchased from Sigma.

Tissue culture flasks, 6 well plates and 96 well plates were obtained from TPP. Annexin V FITC Kit was obtained LY294002 PI3K 阻害剤 from eBioscience Inc.. Real Genomics Total RNA extraction kit and GenomeLab GeXP Start Kit were also procured. Cell culture The human adenocarcinoma breast cancer cell line, MCF 7, and mouse fibroblast cell line, 3T3 were obtained from the American Type and Culture Collection. Cells were cultured in RPMI 1640 supplemented with 10% fetal bovine serum and 1% penicillin and streptomycin, and maintained in humidified incubator at 37 C in atmosphere of 5% CO2. Preparation of EADs The root powder of D. suffruticosa was supplied by Primer Herber Sdn. Bhd.. The plant with voucher specimen number SK1937 11 was deposited in the herbarium of Institute of Bioscience, Universiti Putra Malaysia.

Briefly, 100 g of the powder was soaked in 300 mL of hexane at a ratio of 1,3 with occasional shaking using a rotary shaker for three times at 3,1,1 day interval. The mixture solvent was collected and filtered using Whatman No. 1 filter paper. The residue was dried in an oven at 40 C and subsequently used for successive extraction of dichloromethane followed by ethyl acetate using the same methods. Lastly, filtered ethyl acetate extract was evaporated using a vacuum rotary evapo rator. The yield was weighed and kept at 20 C until required. For subsequent expe riment, the stock of EADs in DMSO was used. The final concentration of DMSO was 0. 33% in all the extracts prepared.