Development of DNA Array for Glioma Expression Profiling by Determination of Genotypes Using of Yeast Assays

本研究旨在建立一种低成本的脑胶质瘤患者个体化诊断方法。我们通过这项研究获得了以下结果。1）我们开发了一种基于酵母的终止密码子测定法，用于检测突变，该测定法易于适用于任何基因（通用终止密码子测定法）（Am J Pathology）。2）我们开发了由1300个基因组成的原始DNA阵列。3）分析了U251 MG、SF 268、SF 295、SF 539、SNB-75和SNB-78等胶质母细胞瘤细胞系的p53、APC和PTEN突变状态及其表达谱。共鉴定出85个与p53突变状态相关的基因。4）研究p53突变对胶质瘤细胞化疗和放疗敏感性的影响。我们发现p53显性负突变（致癌）的生物学意义。我们开发了一个用于胶质瘤表达谱分析的DNA阵列系统。5）我们发现应用特征子集选择算法和神经网络来提取基因表达的特征模式。我们现正就多宗神经胶质瘤的临床个案进行研究。

This study aimed to establish a diagnostic method for personalization of patients with brain gliomas using low-cost DNA array. We obtained the following results through this study.1) We developed a yeast-based stop codon assay to detect mutations which is readily applicable to any genes (universal stop codon assay) (Am J Pathology).2) We developed an original DNA array consisting of 1300 genes. We analyzed 119 cancer cell lines including glioblastoma cells with the array, and obtained successful results.3) We analyzed glioblastoma cell lines including U251MG, SF268, SF295, SF539, SNB-75, and SNB-78, for their p53-, APC- and PTEN-mutational states and their expression profiles. We identified a total of 85 genes that associated with the p53 mutational status.4) We studied effects of p53 mutations on sensitivity of glioma cells to chemotherapeutic agents and radiation. We found a biological significance of p53 dominant negative mutation (Carcinogenesis). We developed a DNA array system for expression profiling of gliomas.5) We found that application of feature subset selection algorithms and neural networks to extract characteristic patterns of gene experssion. We are now undertaking studies on a number of clinical cases of gliomas.