BREAST CANCER MOLECULAR CYTOGENETICS

乳腺癌分子细胞遗传学

• 批准号: 6269283
• 负责人: Frederic M. Waldman
• 金额: $ 23.05万
• 依托单位: CALIFORNIA PACIFIC MED CTR RES INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-04-01 至 1999-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6269283
• 关键词: breast neoplasms; cancer risk; cell population study; cytogenetics; fluorescent dye /probe; gene redundancy; genetic library; genetic markers; human genetic material tag; human subject; immunofluorescence technique; in situ hybridization; molecular oncology; neoplasm /cancer genetics; neoplasm /cancer invasiveness; neoplastic process; nucleic acid hybridization; nucleic acid probes; prognosis

The aim of this project is to characterize cytogenetic changes present in interphase breast tumor cells. Cytogenetic analysis leads to a better understanding of molecular defects in tumor DNA, is an independent prognostic marker for tumor behavior, and will help to pinpoint chromosome regions of particular interest for further molecular analysis. The utility of classical cytogenetics by analysis of metaphase chromosomes has been limited in solid tumors due to the difficulty in preparing adequate tumor metaphases from biopsy tissue. Analysis of tumor cells after short term culture is selective for diploid cells capable of proliferating in restricted culture conditions. Techniques are now available for cytogenetic analysis of interphase tumor cells on a cell by cell basis. Chromosome copy number and major structural aberrations will be detected using fluorescence in situ hybridization with chromosome-specific DNA probes. Copy number of each chromosome will be detected using peri-centromeric repetitive probes specific for chromosomes 1, 7, 11, 133, 16, and 17. These chromosomes have been chosen based on their likely involvement in DNA aberrations. Probe libraries specific for low copy number sequences lining the entire chromosome will be used to "paint" these chromosomes in order to detect translocations or duplications of major chromosome regions. As further probes specific for chromosomal subregions of interest are developed by our collaborators, they will be used to detect amplifications, deletions, and translocations of smaller regions. Similar techniques will be used to characterize to what extent cytogenetically defined subpopulations of tumor cells have proliferative advantage over other subpopulations. Cell proliferative status will be determined by immunofluorescent characterization of bromodeoxyuridine incorporation simultaneously with cytogenetic characterization by fluorescence in situ hybridization. Two and three color analysis will allow comparison of proliferation between populations defined by chromosome copy number or the presence of translocations. The degree of heterogeneity of chromosome abnormalities in tumors can be defined using fluorescence in situ hybridization with chromosome specific probes. Cytogenetic heterogeneity is related to genetic instability and thus to tumor development and progression. The distribution of chromosome abnormalities will be determined using in situ hybridization in tissue sections, and will be compared to tumor architecture and to the distribution of DNA content and antigenic markers. Within the Program Project, cytogenetic information will be correlated with tumor pathology, clinical followup, and related molecular studies.

该项目的目的是表征细胞遗传学变化 间期乳腺肿瘤细胞。 细胞遗传学分析可提供更好的结果 对肿瘤 DNA 分子缺陷的理解是一项独立的研究 肿瘤行为的预后标志物，有助于查明染色体 用于进一步分子分析的特别感兴趣的区域。 实用性 通过中期染色体分析的经典细胞遗传学 由于难以制备足够的肿瘤，因此仅限于实体瘤 来自活检组织的中期。 短期后肿瘤细胞分析 培养物对能够增殖的二倍体细胞具有选择性 培养条件受限。 现在可用于间期肿瘤的细胞遗传学分析 细胞逐个细胞地进行。 染色体拷贝数和主要结构 畸变将使用荧光原位杂交来检测 染色体特异性 DNA 探针。 每条染色体的拷贝数为 使用染色体特异性的围着丝粒重复探针进行检测 1、7、11、133、16 和 17。这些染色体是根据 他们可能与 DNA 畸变有关。 特定于的探针库 整个染色体上的低拷贝数序列将用于 “绘制”这些染色体以检测易位或重复 主要染色体区域。 作为染色体特异性的进一步探针 感兴趣的次区域是由我们的合作者开发的，它们将 用于检测较小片段的扩增、缺失和易位 地区。 类似的技术将用于表征到什么程度 细胞遗传学定义的肿瘤细胞亚群具有增殖能力 相对于其他亚群的优势。 细胞增殖状态将 通过溴脱氧尿苷的免疫荧光表征测定 与细胞遗传学表征同时掺入 荧光原位杂交。 二色和三色分析将 允许比较由染色体定义的群体之间的增殖 拷贝数或易位的存在。 肿瘤中染色体异常的异质性程度可以表示为 使用荧光原位杂交与染色体特异性进行定义 探针。 细胞遗传学异质性与遗传不稳定性有关 从而影响肿瘤的发生和进展。 染色体的分布 将使用组织中的原位杂交来确定异常 切片，并将与肿瘤结构和 DNA含量和抗原标记的分布。 计划内 项目，细胞遗传学信息将与肿瘤病理学相关， 临床随访和相关分子研究。