Apoptosis sensizition of neuroblastoma by altering PI3-K mediated survival signaling

Neuroblastomas are malignant tumors of the central nervous system that usually are found in infants, children, and young adults. These tumors are thought to arise from primitive (undifferentiated) nerve cells left over from the development of the nervous system. Better understanding of the regulatory cell death pathways in neuroectodermal tumors is anticipated to provide new opportunities for design of innovative, molecular targeted therapies. The most important pathway is apoptosis. The aim of this study; to investigate PI3-K signal the impact of mechanism to apoptosis. Beside of them PI3-K is one of the apoptosis related survival transduction pathway proteins. Deregulated PI3-K/Akt signaling often play central role in tumor formation and progression and frequently observed in a variety of human cancer. Thus PI3-K signaling appears to be promising target in the clinical management neuroblastoma patients. Results to be obtained will be allowed especially for cell cycle defined point for developing new targeting therapies.In this study, role of PI3-K/Akt signalling in apoptosis regulation were investigated in neuroblastoma cell lines; SH-EP, lan 5, Kelly. Inhibitors are specific signalling molecules including BGT 226, BEZ 235, BKM 120. As Cytototoxic agent doxorubicin has been used for combine treatment. To find specific proteins; western blotting, to measure the percentage of quantitavie apoptotoic cells nicoletti procedures were applied.As a result of study, differences post treatment between 3 different PI3-K inhibitors, with control group were found significant differences (p<0.05). The neuroblastoma cell lines, have different genetic properties, different responses to inhibitors which were used. Lan 5 cells most sensitive, SH-EP cells were found to be least sensitive cells to this PI3-K inhibitors. SH-EP cells were found to be main target of PI3-K inhibitors. Different concentrations of PI3-K inhibitors as a single agent have been tried in this experiments, it was different response in these cells. There is unsignificant difference between 24 hour, 48 hour and 72 hour other than lan 5 cell line. The apoptosis which occurred in this neuroblastoma cell lines is proved to be caspase dependent apoptosis. Effects of PI3-K inhibitors : BGT 226>BKM 120>BEZ 235. p53 and Bcl-2 genes were investigated in this cycle. For this study should not forget the fact that high concentrations of toxic agents which lead to necrosis other than apoptosis. In this study with nicoletti procedurses, quantitative apoptosis percentage can be performed with the facs analysis. In later stages in this study apoptotic or necrotic cells can be detected with propidium iodide. Method of annexin 5 or with dapi method, apoptotic cells, necrotic cells, live cells can be distinguished. Similar to this study should be try cells of human but not cancer and to compare with this results.