DETECTION OF P53 MUTATIONS IN EWING'S SARCOMA AND RHABDOMYOSARCOMA

尤因氏肉瘤和横纹肌肉瘤中 P53 突变的检测

• 批准号: 3796579
• 负责人: M TSOKOS
• 金额: --
• 依托单位: DIVISION OF CANCER BIOLOGY AND DIAGNOSIS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3796579
• 关键词: Ewing's tumor; clone cells; family genetics; gel electrophoresis; gene mutation; human tissue; molecular oncology; neoplasm /cancer genetics; pediatric neoplasm /cancer; polymerase chain reaction; rhabdomyosarcoma; structural genes; tissue /cell preparation; tumor suppressor genes

p53 mutations have been detected in many tumors and interpreted as a late event in tumorigenesis, probably involved in tumor progression rather than tumor development. Recent studies have demonstrated the presence of germline p53 mutations in some Li-Fraumeni pedigrees and hence a role of p53 mutations in hereditary susceptibility to human cancer. Rhabdomyosarcoma, a common soft tissue sarcoma in the families with the Li- Fraumeni syndrome, was found to have p53 mutations. However, the frequency of such mutations in pediatric tumors in general remains unknown. We will study the presence of p53 mutations in established Ewing's sarcoma cell lines in comparison with existing data from rhabdomyosarcoma cell lines. In addition, p53 mutations will be studied in paraffin embedded tissues from rhabdomyosarcoma tumors to investigate the validity of paraffin embedded tissues for such studies. The detection of mutations will be performed on PCR fragments spanning exons 4, 5, 6, 7, and 8 of the p53 gene. Two methods will be compared: the standard single stranded conformational polymorphism (SSCP) method, and a non-radioactive method, based on conformational differences in double stranded molecules (heteroduplex analysis) and using non-denaturing electrophoresis in mutation detection enhancement (MDE) gels.

在许多肿瘤中都检测到p53突变，并解释为迟到 肿瘤发生中的事件，可能参与肿瘤进展而不是 肿瘤发育。 最近的研究表明存在 某些Li-Fraumeni的种系P53突变，因此 遗传性易感性的p53突变。 横纹肌肉瘤，一种常见的软组织肉瘤 发现Fraumeni综合征有P53突变。 但是，频率 一般来说，小儿肿瘤中的这种突变仍然未知。 我们将 研究已建立的Ewing肉瘤细胞中p53突变的存在 与来自横纹肌肉瘤细胞系的现有数据相比。 此外，将在石蜡嵌入组织中研究p53突变 从横纹肌肉瘤肿瘤来研究石蜡的有效性 嵌入的组织进行此类研究。 突变的检测将是 在跨越外显子4、5、6、7和8的PCR片段上执行P53 基因。 将比较两种方法：标准单链 构象多态性（SSCP）方法和非放射性方法， 基于双链分子的构象差异 （杂化分析）并在中使用非降低电泳 突变检测增强（MDE）凝胶。