Characterization of the Ews/Fli-1 Chimeric Gene

描述(申请人提供)：神经母细胞瘤(NB)和尤文氏肉瘤/外周原始神经外胚层肿瘤(ES/PNET)是儿童神经脊来源的癌症。有人认为NBS交感神经谱系与ES/PNETs副交感神经谱系的分化程序是以线性方式进行的。对NB细胞系和ES/PNET细胞系的体细胞杂交显示，NB特异性标记被抑制，而将ES/PNETs中特有的EWS/FLI-1嵌合基因导入NB也显示了类似的结果。本项目将利用蛋白质印迹分析检测P53和pRb的表达，免疫荧光检测P53的定位，蛋白半衰期实验检测P53的稳定性，研究EWS/Fli-1‘S在细胞周期调控中的作用。本项目还将利用芯片阵列技术研究EWS/FLI-1癌基因的机制和途径。

DESCRIPTION (provided by applicant): Neuroblastomas (NB) and Ewing's Sarcoma/peripheral Primitive Neuro-ectodermal Tumors (ES/PNET) are pediatric cancers of neural crest orgin. It has been suggested that the control of the differentiation program o f the sympathetic lineage of NBs versus the parasympathetic lineage of ES/PNETs proceed in a linear fashion. Somatic cell hybridization of NB cell lines and ES/PNET cell lines show that NB-specific markers are repressed, and the transfection of NB with the Ews/FLI-1 chimeric gene characteriztically found in ES/PNETs show similar results. This project will characterize the Ews/FLI-1's role in cell cycle control utilizing western blot analysis to monitor expression of p53 and pRb, immunofluorescence to monitor p53 localization and protein half life experiments to monitor p53 stability. This project will also utilize microchip array techinology to study the mechanism and pathway of the Ews/FLI-1 oncogene.