The functional implications of Ras hyper activation TNF stimulation, Elevated angiogenesis and increased breast tumor cell dissemination to lymph nodes The results obtained thus far in this study indicate that the cooperative activities of TNF with RasG12V or with WT Ras lead to additive elevation in the release of CXCL8 by the tumor cells. Similarly, many other pro cancerous factors may further info be induced in TNF Ras stimulated cells. The outcome of such a process, if taking place in vivo in malignancies with high TNF expression as is the case in breast cancer may be high production of pro tumorigenic factors by the tumor cells, including Inhibitors,Modulators,Libraries angiogenic ones. To examine whether such a general increase in pro tumoral and angiogenic factors indeed leads to increased angiogenesis, we used the in vivo analysis of chorioallan toic membrane assay.

In this test, multiple pa rameters of angiogenesis are Inhibitors,Modulators,Libraries affected by angiogenic factors, including length and thickness of blood vessels and their sprouting. Due to its multi parametric nature, Inhibitors,Modulators,Libraries to the high content of vessels in the embryo and to em bryo heterogeneity, the results of the CAM assay often show variability between individual samples within the same group, thus, the CAM assay could Inhibitors,Modulators,Libraries clearly define differences between two extreme conditions, but its sensitivity could not de termine interim effects that may have been obtained by other combinations that are less effective in inducing an giogenic and pro tumoral factors.

To comply with this limitation, and in line with our Inhibitors,Modulators,Libraries interest in determining the overall effects induced by multiple angiogenic factors that could have been promoted by the most potent process of TNF stimulation of WT Ras expressing cells, we tested CM from the two most relevant stimula tory extreme conditions, CM of WT Ras expressing tumor cells that were stimulated by TNF. CM of control vector expressing tumor cells that were not stimulated by the cytokine. The results indicate that CM derived from TNF stimulated WT Ras expressing tu mor cells induced significantly stronger angio genic effects compared to control cells. In parallel, we asked what is the impact of combined TNF stimulation and Ras hyper activation on tumor growth and metastasis. MCF 7 cells were documented as cells with relatively low malignancy potential, and with very weak invasive and metastasizing capacities. However, published studies by Weinberg and his colleagues have shown that selleck under specific conditions, MCF 7 cells that express oncogenic Ras can form metas tases. Thus, to allow for metastatic dissemination in our study, we followed on these observations and used RasG12V expressing MCF 7 cells, compared to cells transfected with control vector.