, 2006). Although the expression of activated AKT1 accelerates HER-2/NEU-driven breast tumor formation, the tumors that developed were highly differentiated, poorly invasive, and rarely metastasized ( Hutchinson et al., 2004). AKT1 also plays a prominent role in tumor angiogenesis. Normal endothelial cells with sustained activation of AKT1 develop the complex structural and functional abnormalities that are characteristic of tumor blood vessels ( Phung et al., 2006). These results

reinforce the idea that an understanding of cell- and tissue-specific NVP-BKM120 manufacturer signaling pathways is critical for evaluating the implications of activated upstream signaling molecules on complex phenotypic effects. Until now, despite large research efforts in targeting tumor metastasis, no progress has been achieved in efficiently preventing metastasis (Christofori, 2006). This might be

due to the mechanisms involved in cell migration, which can be reprogrammed, thus allowing the cells to maintain their invasive properties via morphological and functional de-differentiation. Natural products have been remarkable source for new anticancer drugs. Biflorin (Fig. 1), an οrtho–naphthoquinone, can be isolated from the roots of Capraria biflora L. (Schrophulariaceae),a perennial shrub that was originally found in the Antilles and South America ( Acosta et al., 2003). This quinone has been shown to have anticancer properties in vitro and in vivo, increasing the survival of mice with melanoma tumors, without diminishing the tumor size ( Vasconcellos et al., 2005, Vasconcellos et

al., 2007 and Vasconcellos http://www.selleckchem.com/products/VX-765.html et al., 2011). As such, the aim of this work is to investigate the role of biflorin in MDA-MB-435, an invasive melanoma cancer cell, in vitro. The MDA-MB-435 (human melanoma), MCF-10A (normal human breast) and melan-A (normal mouse melanocyties) cell lines were obtained from American Type Culture Collection (ATCC). All the cell lines were cultured according to ATCC recommendations. The following reagents were used: mouse monoclonal anti-N-cadherin AB-2 (Cell Signaling), mouse monoclonal anti-β-actin (Cell Signaling), and Super Signal West Pico Chemiluminescent kit (Pierce). Etoposide, dimethyl sulfoxide, paraformaldehyde, and crystal violet were purchased from Sigma–Aldrich. Alamar Blue was Isotretinoin purchase from Invitrogen. The invasion plates were obtained from Corning. Cell viability assays. MDA-MB-435 cells were seeded in 96-well plates at a density of 104 cells per well. They were treated with biflorin, and the Alamar BlueTM assay was performed (Ahmed et al., 1994) after 8, 12, and 24 h. After the cells were allowed to attach for 24 h, biflorin (0.1, 0.5, 1, 5 and 10 μM) was dissolved in dimethyl sulfoxide (DMSO) and added to each well, and the cells were incubated for 8, 12 and 24 h. Etoposide (10, 20 and 50 μM) was used as positive control. Control groups received the same amount of DMSO (0.1%).