Molecular Studies of Esophageal Adenocarcinoma

描述（由申请人提供）：本次更新申请的目的是鉴定食管癌中重要的和潜在的新型扩增癌症相关基因，食管癌是一种发病率正在增加的致命癌症。基因组扩增的鉴定，以及由此产生的重要基因在这些肿瘤中的过表达，将为了解与该疾病的发生和进展相关的分子事件提供重要的见解。这项工作可能为早期检测提供新的遗传工具，并有可能为治疗干预确定新的途径。我们使用高灵敏度的定量二维基因组扫描（2D凝胶）方法和使用寡核苷酸阵列的基因表达谱，在食管腺癌中发现了许多潜在的新的基因组扩增事件。2d凝胶技术涉及对来自个体患者正常和肿瘤组织之间的基因组功能成分的数千个基因组限制性片段进行比较分析。扩增子中包含的扩增和过表达基因被鉴定和表征，我们已经成功地证明了这一点。利用定义明确的染色体标记，定量PCR检测和大量正常组织-腺癌配对样本的DNA，我们已经确定了一些特定扩增子的“最小共同扩增区域”，用于立即分析，其他扩增子将被确定。重要的扩增和过度表达的癌症相关基因将被定义。潜在的候选基因可能包括已知基因，未表征的表达序列标签，或从最小区域内的基因组克隆中分离的基因。新基因的癌症相关性将通过功能分析进行检验，包括转染到永生化细胞中，分析对细胞生长、侵袭性、锚定独立性、细胞凋亡的影响，并在选定的情况下在小鼠中形成肿瘤。在体外研究中，使用含有扩增基因的腺癌细胞或转染后含有过表达基因的腺癌细胞，将被用于潜在的靶向治疗，从而为鉴定特定扩增事件提供临床相关性。这些研究将确定在食管腺癌中扩增的特定基因的身份和频率，并对这种重要癌症的发生和/或进展的机制提供更深入的了解。

DESCRIPTION (provided by applicant): The goals of this renewal application are to identify important and potentially novel amplified cancer-related genes in esophageal adenocarcinomas, a deadly cancer that is increasing in incidence. The identification of genomic amplifications, and the resulting important genes over-expressed in these tumors, will provide significant insight into the molecular events associated with the development and progression of this disease. This work may provide new genetic tools for early detection, and potentially for identifying new avenues for therapeutic intervention. We have identified a number of potentially novel genomic amplification events in esophageal adenocarcinomas using the highly sensitive method of quantitative two-dimensional genomic scanning (2D gels) and using gene expression profiling with oligonucleotide arrays. The 2D-gel technology involves the comparative analysis of several thousand genomic restriction fragments derived from functional components of the genome between the normal and tumor tissue from an individual patient. The amplified and over-expressed genes encompassed in the amplicons are identified and characterized as we have successfully demonstrated. Utilizing well-defined chromosomal markers, a quantitative PCR assay and the DNA from large numbers of normal tissue-adenocarcinoma paired samples, we have identified the "minimal region of common amplification" for a number of specific amplicons for immediate analysis and other amplicons will be determined. The important amplified and over-expressed cancer-related genes and will be defined. Potential candidates may include known genes, uncharacterized expressed sequence tags, or genes isolated from genomic clones that map within the minimal region. The cancer-relatedness of novel genes will be examined using functional assays involving transfection into immortalized cells and the analysis of the effects upon cell growth, invasiveness, anchorage-independence, apoptosis and in selected cases tumor formation in mice. In vitro studies using adenocarcinoma cells containing amplified genes of interest of containing overexpressed genes as the result of transfection will be examined for potential targeted therapy and thus providing clinical relevance to identification of specific amplification events. These studies will determine both the identity and frequency of specific genes amplified in esophageal adenocarcinomas and also provide a greater insight into the mechanisms underlying the development and/or progression of this important cancer.