STROMAL INFLUENCES ON BREAST CANCER PROGRESSION

间质对乳腺癌进展的影响

• 批准号: 2008719
• 负责人: SANDRA W MCLESKEY
• 金额: $ 11.14万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-01-01 至 1999-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2008719
• 关键词: MCF7 cell; RNase protection assay; angiogenesis; angiogenesis factor; biomarker; breast neoplasms; fibroblast growth factor; genetic transcription; immunocytochemistry; in situ hybridization; microcirculation; neoplasm /cancer blood supply; neoplasm /cancer pharmacology; neoplastic process; pentosan; polymerase chain reaction; receptor expression; stainings; stromal cells; tamoxifen; transfection; vascular endothelium

Transfection of MCF-7 breast carcinoma cells with fibroblast growth factor 4 (FGF-4), an angiogenic growth factor, produces a dramatic change in tumor phenotype when the cells are injected into the mammary fat pads of ovariectomized nude mice. While the parental cells are estrogen- dependent for tumor growth, poorly invasive, and rarely metastatic, the transfectants produce large, progressively growing tumors which are invasive, frequently metastatic and growth-stimulated by tamoxifen, an estrogen antagonist. This in vivo behavior of the transfectants is in contrast to their in vitro behavior, which is not substantially different from the parental MCF-7 cells (specifically, the transfectants are not growth-stimulated by tamoxifen in vitro). Thus, it would seem likely that at least some portion of the change in in vivo phenotype is due to an interaction between the transfected gene product and stromal components of the tumor. Since the transfected gene product is a known angiogenic growth factor, it seems likely that increased angiogenesis in tumors produced by-the transfected cells is at least partially responsible for the change in in vivo phenotype produced by the transfection. A number of FGF receptor (FGFR) genes have been identified which may each encode multiple receptors via alternative mRNA splicing. Evidence is emerging which attributes specific responses or affinity for specific FGF ligands to specific FGFR isoforms. Specific FGFRs in tumors produced by transfected cells may respond to the FGF-4 ligand and/or be up- or down- regulated. Evidence is also accumulating that endothelial cells from specific tissues differ, and that endothelial cells from tumors are different from those in normal tissues. Cultured endothelial cells are quite plastic in their gene expression and behavior depending on their passage and growth conditions. Therefore, attempts to study tumor angiogenesis in vitro must use appropriate endothelial cells under appropriate conditions. This project seeks to relate changes in the process of angiogenesis in tumors produced by parental and FGF-4 transfected MCF-7 cells to the change in in vivo behavior. We will relate microvessel density and total vascularity to tumor size in tumors produced by parental and transfected cells. We will describe the process of angiogenesis in tumors produced by parental and transfected cells by techniques which identify proliferating or immature endothelial cells in tumor sections obtained at very early and later time points after injection of tumor cells. The project will also seek to characterize specific in situ FGFR expression by endothelial and other cells contained within the tumor. We will isolate endothelial cells from tumors produced by parental or FGF-4 transfected MCF-7 cells as well as endothelial cells from normal mammary fat pads. Growth assays in which quiescent cells are stimulated to invade a 3-dimensional matrix will be done utilizing very early passage tumor-derived and normal endothelial cells to identify differences in behavior between the three types of endothelial cells. Tumor-derived and mammary fat pad endothelial cells will be subjected to differential display PCR in a search for genes which are preferentially expressed in tumors produced by FGF-4 transfected MCF-7 cells. Transcription of such genes might be an important downstream event of FGFR stimulation and the protein products of such genes might be important in determining the phenotype of the tumor.

成纤维细胞生长的MCF-7乳腺癌细胞的转基因研究 4因子(FGF-4)是一种血管生成生长因子，它能产生巨大的变化 当细胞被注射到乳房脂肪垫中时，肿瘤表型 去卵巢的裸鼠。当亲代细胞是雌激素时- 依赖于肿瘤生长，侵袭性差，很少转移， 转染者产生巨大的、渐进性生长的肿瘤 他莫昔芬，一种侵袭性的，经常转移和刺激生长的 雌激素拮抗剂。这种转染者的体内行为是在 与它们的体外行为形成对比，这并没有本质上的区别 从亲本MCF-7细胞(具体地说，转染体不是 生长-由他莫昔芬在体外刺激)。因此，看起来很可能 体内表型的改变至少有一部分是由于 转基因产物与基质成分的相互作用 肿瘤的组织结构。由于所导入的基因产物是已知的血管生成 生长因子，似乎有可能增加肿瘤的血管生成 产生于--转基因细胞至少部分负责 转基因引起的体内表型的变化。一个数字 的成纤维细胞生长因子受体(FGFR)基因已被鉴定，每个基因都可能编码 多个受体通过选择性的信使核糖核酸剪接。证据正在浮现 哪些属性为特定的成纤维细胞生长因子配体的特定反应或亲和力 与特定的FGFR亚型结合。在肿瘤中产生的特异性FGFRs 转基因细胞可能对成纤维细胞生长因子-4配体产生反应和/或上调-或下调- 受监管的。越来越多的证据表明，血管内皮细胞 特定的组织不同，肿瘤的内皮细胞是 与正常组织中的不同。培养的内皮细胞 它们的基因表达和行为相当可塑性，这取决于它们 传代和生长条件。因此，试图研究肿瘤 体外血管生成必须使用适当的血管内皮细胞 适当的条件。这个项目寻求将 亲本和成纤维细胞生长因子-4诱导的肿瘤血管生成过程 转染后的MCF-7细胞在体内行为的改变。我们会 肿瘤微血管密度和总血管密度与肿瘤大小的关系 由亲本细胞和转基因细胞产生。我们将描述这一过程 亲本细胞和转基因细胞产生的肿瘤血管生成的研究 识别血管内皮细胞增殖或未成熟的技术 肿瘤切片在很早和很晚的时间点分别在 注射肿瘤细胞。该项目还将寻求将 血管内皮细胞和其他含有FGFR的细胞特异性原位表达 在肿瘤内。我们将从产生的肿瘤中分离内皮细胞 经亲本或成纤维细胞生长因子4基因转染的MCF-7细胞及内皮细胞 来自正常的乳房脂肪垫。生长试验，其中静止细胞是 被刺激入侵一个3维矩阵将使用非常 早期传代肿瘤来源的内皮细胞和正常内皮细胞的鉴定 三种类型的内皮细胞之间的行为差异。 肿瘤来源的内皮细胞和乳腺脂肪垫内皮细胞将受到 差异显示聚合酶链式反应在基因搜索中的应用 在转染成纤维细胞生长因子-4的MCF-7细胞产生的肿瘤中表达。 这些基因的转录可能是一个重要的下游事件 FGFR刺激和这些基因的蛋白产物可能是 对于确定肿瘤的表型很重要。