CORE--MUTATION DETECTION & CHARACTERIZATION OF TUMOR SUPPRESSOR GENES

该核心的主要功能是检测肉瘤激酶中的p53突变并对其进行功能表征。这些分析对项目1、4和5的目标至关重要。Strong博士（项目1）使用从该核心中获得的数据来表征儿童肉瘤患者运动中的过度癌症风险，以评估p53生殖系突变对儿童肉瘤的相对贡献，并确定新发突变的父亲来源。此外，由于我们发现了非p53癌症倾向的激酶，她也将描述这些激酶的癌症风险。她进行这些分析的能力取决于确定哪些个体携带p53生殖系突变。该核心获得的数据对于Lozano博士的目标（项目4）也是必不可少的，即在非p53癌症易感性激酶中定位负责癌症易感性的基因座或基因座，因为额外的家族和个体的鉴定加强了连锁分析所需的能力。为了确定与p53突变相关的独特的定量微卫星不稳定表型（项目5，Siciliano博士），显然有必要鉴定具有生殖系p53突变的患者。该核心的具体目标是：1）对来自软组织肉瘤和骨肉瘤组群的先证者的p53编码外显子和侧翼剪接点进行测序，以鉴定生殖系p53突变的存在或不存在2）通过Southern分析来检查其中p53突变未被上述（1）鉴定的那些样品，以排除p53等位基因之一的可能的总体缺失3）检测有无p53突变的LFS患者肿瘤和永生化成纤维细胞中的洛缺失4）发展SSCP和阿索方法，以筛选特异性p53突变的家族成员，并筛选非p53激酶中的其他已知抑癌基因5）使用酵母功能测定和组织培养细胞中的瞬时转染实验进行p53突变的功能分析，突变之间的差异。这个核心的功能最终将发展到包括其他基因或易患癌症的基因的突变分析（见项目4和核心E）。

The main functions of this core are the detection and functional characterization of p53 mutation in sarcoma kindreds. These analyses are essential for the aims of projects 1, 4, and 5. Dr. Strong (project 1) uses the data obtained from this core to characterize excess cancer risk in kindreds of childhood sarcoma patients, to assess the relative contribution of p53 germline mutations to childhood sarcoma, and to identify paternal origin of de novo mutations. Additionally, since our discovery of non-p53 cancer prone kindreds, she will characterize cancer risk in these kindreds as well. Her ability to do these analyses is dependents of establishing which individuals carry p53 germline mutations. The data obtained by this core are also essential for Dr. Lozano's aim (project 4) to map the locus or loci responsible for cancer predisposition in non p53 cancer prone kindreds since the identification of additional families and individuals strengthens the power needed for linkage analysis. To determiner where there is a distinct quantitative microsatellite instability phenotype associated with p53 mutations (project 5, Dr. Siciliano), it is clearly essential to identify the patients with germline p53 mutations. The specific aims of this core are: 1) to sequence the p53 coding exons and flanking splice junctions from the probands of the soft tissue sarcoma and osteosarcoma cohorts to identify the presence or absence of germline p53 mutations 2) to examine those samples in which a p53 mutation is not identified by (1) above by southern analysis to exclude possible gross deletions of one of the p53 alleles 3) to examine LOH in tumors and immortalized fibroblasts from LFS patients with and without p53 mutations 4) to develop SSCP and ASO methods to screen family members upon identification of the specific p53 mutation and to screen o6ther known tumor suppressor genes in non p53 kindreds 5) to perform functional analyses of p53 mutations using the yeast functional assay and transient transfection experiments in tissue culture cells to document functional differences between mutations. The function of this core will eventually evolve to include mutational analyses of the other gene or genes that predispose to cancer (see project 4 and core E).