Tissue Microarray Design and Development for Novel Gene and Gene Product Screeni

确定在肿瘤细胞中重要的基因和基因产物是国家癌症研究所的高度优先目标。为了确定感兴趣的基因是否在人类恶性肿瘤中表达，研究人员必须能够评估人类肿瘤的实际组织样本。不幸的是，获取大量特征良好的人体组织样本是困难的，而且为了进行免疫组织化学或原位杂交，制备肿瘤玻片的费用很高。作为这一问题的部分解决方案，病理学家开始将多个组织样本组合在一个组织块中，以减少组织学成本和进行特殊研究所涉及的时间和精力。这项技术已经改进到可以将数百个组织样本以网格方式放置在单个石蜡组织块中。NHGRI的研究人员(Kohenen、Kallioniemi等)在《科学》杂志上发表了一篇开创性的论文，展示了大规模组织微阵列的用途。<br><br>1999年秋季，NCI主办了一次组织微阵列研讨会，建立了一个以病理学为基础的组织微阵列核心设施。该项目是2001年旁路预算计划中确定癌细胞特征的非凡机会的直接副产品。第一年的目标是在NHGRI类似设施的基础上建立一个核心微阵列生产设施，并创建第一个在全国范围内分发的混合肿瘤微阵列。核心的最初产品是一个混合肿瘤块，其中包含最常见的上皮性恶性肿瘤(乳腺、结肠、肺、前列腺和卵巢)的代表性样本以及黑色素瘤、胶质瘤和淋巴瘤的样本。选择正常组织和标准细胞系也包括在内，使组织斑点总数达到500个。匿名的人类肿瘤样本是通过CHTN获得的，该组织还将在玻璃载玻片上向壁内和壁外科学界的研究人员分发阵列。在第一个生产年度(2001财年)准备了四套多肿瘤阵列，总计3000多张幻灯片用于分发。在这个项目的当前阶段，我们主要是在肿瘤特定项目上与内部调查人员合作，既建立阵列，也染色和分析病理。在过去的一年(2005财年)，我们已经建立了NCI60细胞系和来自这些细胞系的异种移植细胞系的阵列，这些细胞系包括儿童肿瘤、乳腺癌、非小细胞肺癌、黑色素瘤和普通肿瘤阵列。除了幻灯片生产，核心设施还从事技术开发，从基本的组织学程序到阵列使用，再到阵列的大规模成像。我们已经购买了一台自动阵列机，并在自动阵列机和自动成像系统的进一步技术开发方面进行了合作。

Identifying genes and gene products that are important in tumor cells is a high priority goal of the National Cancer Institute. In order to determine whether a gene of interest is expressed in human malignancy, investigators must be able to evaluate actual tissue samples of human tumors. Unfortunately, access to large numbers of well-characterized human tissue samples is difficult, and there is a large expense for the preparation of glass slides from tumor in order to perform immunohistochemistry or in-situ hybridization. As a partial solution to this problem, pathologists began to combine multiple tissue samples in one tissue block, in order to reduce the histology costs and the time and effort involved to perform the special studies. This technique has been refined to the point where hundreds of tissue samples can be placed in a grid arrangement in a single paraffin tissue block. Investigators in the NHGRI (Kohenen, Kallioniemi and others) demonstrated the utility of large scale tissue microarrays in a seminal paper published in Science.<BR><BR>Following a tissue microarray workshop hosted by NCI in Fall, 1999, a pathology-based tissue microarray core facility was established. This project is a direct outgrowth of the Extraordinary Opportunity to Define the Signatures of Cancer Cells identified in the 2001 Bypass Budget plan. The goals of the first year were to establish a core microarray production facility based on a similar facility in the NHGRI and to create the first mixed tumor microarrays for nationwide distribution. The initial product of the core was a mixed tumor block containing representative samples of the most common epithelial malignancies (breast, colon, lung, prostate, and ovary) as well as samples of melanoma, glioma and lymphoma. A selection of normal tissue and standard cell lines was also included to bring the total number of tissue spots to 500. Anonymized human tumor samples are obtained through the CHTN, and this organization will also handle distribution of arrays on glass slides to investigators in the intramural and extramural scientific community. Four sets of multi-tumor arrays were prepared in the first production year (FY2001), totaling more than 3000 slides for distribution. In the current phase of this project we are collaborating with mainly intramural investigators on tumor specific projects, both building arrays as well as staining and analyzing the pathology. This past year (FY2005) we have built arrays of the NCI60 cell lines and xenografts from those cell lines, pediatric tumors, breast carcinoma, non-small cell lung carcinoma, melanoma and general tumor arrays. In addition to slide production, the core facility has been engaged in technology development, from the basic histology procedures to array use to large-scale imaging of arrays. We have acquired an automated arrayer and have worked collaboratively on further technology development of the automated arrayer and an automated imaging system.