DEFINITION OF ENVIRONMENT IN HUMAN BREAST CANCER

人类乳腺癌环境的定义

• 批准号: 2107467
• 负责人: MINA BISSELL
• 金额: $ 18.1万
• 依托单位: UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-06-01 至 1998-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2107467
• 关键词: athymic mouse; basement membrane; biomarker; breast neoplasms; cadherins; cell cell interaction; cell growth regulation; cell type; cellular oncology; cellular polarity; disease /disorder model; extracellular matrix; human tissue; integrins; mammary epithelium; molecular oncology; tissue /cell culture; tumor suppressor genes

DESCRIPTION (Applicant's description) It is increasingly apparent that both the normal, differentiated state, and the manifestation of the malignant phenotype in the epithelia, are dependent on the nature and the integrity of the surrounding stroma and the extracellu- lar matrix (ECM). While these phenomena are relatively well studied in animal models, the definition and the role of the "microenvironment" in the human tissues has lagged behind. My coworkers and I have established culture systems for the cultivation of functional epithelial cells from the rodent mammary gland in which they are able to recreate "lactating" alveoli in the culture dish. As such, we have unraveled some unexpected aspects of microenvironmental control. Together with the Co-Principal Investigators, we have applied these methods to establish a rapid and versatile assay for distinguishing normal and malignant human breast epithelial cells in a 3- dimensional organotypic assay. Our main collaborator (OWP) and his coworkers have isolated and characterized one of a very few "diploid" human cell lines (HMT3522) that in our 3-D assay behaves like normal cells from reduction mammoplasties. During passage in culture, these cells become aneuploid and after 205 passages become tumorigenic in the nude mice, and at this stage behave like the carcinomas in our assay. Passages of this line, as well as primary luminal cells from reduction mammoplasty, will be used for functional studies outlined here. We will choose 5 passages between early, intermediate and late stages of HMT3522, with the first being "normal" by the criteria of our assay, and the last, malignant by the criteria of tumor formation in the nude mice. The aims are to understand, in a well defined culture model which mimics the behavior or the cells in vivo, the influence of stromal cells (primary or secondary) on the function of the epithelial cells and vice versa. We have recently developed techniques for isolation of the major cell types from the human breast stroma. We will further characterize these cells and use them together with cells from primary tumors in 3-D cultures to explore the origin and the nature of the "stromal reaction". We have defined a number of markers to distinguish normal (reduction mammoplasty as well as HMT 3522) and malignant epithelial cells as a result of cell-ECM interactions. These include the loss of ability to form TDLU (terminal duct lobular units), loss of polarity (as measured by the localization of sialomucin and expression of E and P cadherins), changes in integrins, and components of the basement membrane, and importantly, the inability to growth arrest in 3-D. We will supplement the list with additional markers including known oncogenes and tumor suppressor genes reported to change in breast cancer, as well as assays for active TGFbeta and TGFalpha to generate an informative panel of markers. These will be used to measure gene expression and to monitor the progression to tumorigenicity after different combinations of "homotypic and heterotypic" interactions between the stromal and the epithelial cells. The long range goals are a) to define, in a physiological microenvironment, accurate and early markers for changes to the malignant phenotype as a result of stromal- epithelial interactions and b) to pave the way to understanding the detailed mechanism by which the microenvironment regulates function in normal and malignant breast.

描述（申请人的描述） 越来越明显的是，无论是正常的还是分化的 状态以及恶性表型的表现 上皮细胞，依赖于性质和完整性 周围基质和细胞外基质（ECM）。 虽然这些 现象在动物模型中得到了相对充分的研究，定义 而“微环境”在人体组织中的作用 落后了。 我和我的同事已经建立了文化 用于培养功能性上皮细胞的系统 啮齿动物的乳腺，它们能够在其中重建“哺乳期” 培养皿中的肺泡。 因此，我们已经解开了一些 微环境控制的意外方面。 与 联合首席研究员，我们已将这些方法应用于 建立一种快速、通用的检测方法来区分正常和 3维恶性人乳腺上皮细胞 器官型测定。 我们的主要合作者（OWP）和他的同事已经 分离并表征极少数“二倍体”人类细胞系之一 (HMT3522) 在我们的 3-D 检测中，其行为类似于来自 乳房缩小整形术。 在培养过程中，这些细胞 成为非整倍体，并且在 205 次传代后成为致瘤性的 裸鼠，在这个阶段的行为就像我们体内的癌症一样 化验。 该系的传代以及来自的原代管腔细胞 缩小乳房成形术，将用于概述的功能研究 这里。 我们会在早、中、晚之间选择5个段落 HMT3522 的各个阶段，根据以下标准，第一个阶段为“正常” 我们的检测，最后一个，根据肿瘤形成的标准，是恶性的 在裸鼠中。 目的是在明确定义的情况下理解 模拟体内细胞行为的培养模型， 基质细胞（初级或次级）对功能的影响 上皮细胞，反之亦然。 我们最近开发了 从人体中分离主要细胞类型的技术 乳腺基质。 我们将进一步表征这些细胞并使用它们 与 3D 培养中的原发性肿瘤细胞一起探索 “基质反应”的起源和性质。 我们定义了一个 区分正常的标记数量（缩乳术以及 如 HMT 3522）和细胞 ECM 导致的恶性上皮细胞 互动。 其中包括失去形成 TDLU 的能力 （末端导管小叶单位），极性丧失（通过测量 唾液粘蛋白​​的定位以及 E 和 P 钙粘蛋白的表达）， 整合素和基底膜成分的变化，以及 重要的是，无法在 3-D 中停止生长。 我们将 用包括已知癌基因在内的其他标记补充列表 据报道，肿瘤抑制基因也会在乳腺癌中发生变化 作为活性 TGFbeta 和 TGFalpha 的测定，以生成信息丰富的 标记面板。 这些将用于测量基因表达和 监测不同后的致瘤性进展 之间“同型和异型”相互作用的组合 基质细胞和上皮细胞。 长期目标是 a) 在生理微环境中准确且早期地定义 间质导致的恶性表型变化的标志物 上皮相互作用和b）为理解铺平道路 微环境调节功能的详细机制 在正常和恶性乳房中。