Xenobiotics and Breast Epithelial Cell-ECM Signaling

异生素和乳腺上皮细胞 ECM 信号转导

• 批准号: 6518191
• 负责人: RAYMOND F NOVAK
• 金额: $ 26.08万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-05-03 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6518191
• 关键词: apoptosis; breast neoplasms; cell adhesion; cell adhesion molecules; cell cell interaction; cell cycle; cell cycle proteins; cell growth regulation; cell line; chemical carcinogen; chemical carcinogenesis; dichlorodiphenyltrichloroethane; dimethylbenzanthracene; environment related neoplasm /cancer; environmental toxicology; extracellular matrix; gene expression; integrins; mammary epithelium; microarray technology; mirex /kepone; phorbols; phosphatidylinositol 3 kinase

DESCRIPTION: (Adapted from the Applicant's Abstract): considerable controversy exists regarding the association of xenobiotics with breast cancer. Cell-ECM Extracellular Matrix) interactions are critical to cellular morphology, and alterations in cell morphology alter cell unction and gene transcription. Cells are linked to the ECM via integrin-containing focal adhesions. Other types of structures that link cells to each other or to the ECM include adherens junctions and desmosomes. Because correct issue architecture and cell morphology are critical for cellular homeostasis, we have examined whether xenobiotics disrupt cell-ECM interactions in a series of breast epithelial cell lines that represent a model of human proliferative breast disease (PBD), a risk factor for breast cancer. Preliminary results show that cells cultured on the ECM Matrigel adopt a three-dimensional morphology reminiscent of breast architecture in vivo and display differential sensitivity to Fas-mediated apoptosis. Organochlorines disrupt the levels and association of adhesion molecules associated with cell-ECM and cell-cell communication. Treatment of cells with p,p'-DDT results in increased delta-catenin levels relative to controls, whereas Kepone decreases alpha-catenin expression 2-fold and disrupts desmosomes relative to controls. The hypothesis of this research is that xenobiotics alter the levels, localization and association of cell adhesion molecules critical to the formation of focal adhesions, desmosomes and adherens junctions, which ultimately affect intracellular signaling pathways (e.g. PI 3-kinaseIAkt kinase), gene expression, cell cycle regulation, proliferation and apoptosis. We will employ organochlorine (p,p'-DDT, o,p'-DDT and Kepone) and non-organochlorine xenobiotics (DMBA, TPA, PhIP) in this proposal. Thus, the specific aims of this research, employing breast epithelial cells cultured on Matrigel, are: 1) To determine xenobiotic effects on the distribution and association of cell adhesion molecules critical to normal cell-ECM interactions and focal adhesions, adherens junctions or desmosomes. 2) To examine xenobiotic effects on PI 3-kinase/Akt signaling. 3) To determine xenobiotic effects on the cell cycle and cell cycle regulatory components (pRB, E2F, p53, cyclins and cyclin dependent kinases (cdk)). 4) To examine whether xenobiotics alter Fas-mediated apoptosis. 5) To assess whether xenobiotics at concentrations relevant to tissue levels in vivo, alter global gene expression using microarray analysis. The results of this proposal will provide novel information regarding chemical-mediated effects in breast epithelial cells and seminal data on fundamental cellular changes in human proliferative breast disease.

描述：（改编自申请人摘要）：相当大的争议 存在关于异生物质与乳腺癌的关联。细胞外基质 细胞外基质）相互作用对细胞形态至关重要， 细胞形态的改变改变细胞功能和基因转录。细胞 通过含整合素的粘着斑与ECM连接。其他类型的 将细胞彼此连接或连接到ECM的结构包括粘附分子 连接和桥粒。因为正确的问题架构和单元 形态对于细胞内稳态至关重要，我们已经检查了是否 外源性物质破坏一系列乳腺上皮细胞中细胞-ECM相互作用 代表人增殖性乳腺疾病（PBD）模型的线， 乳腺癌的危险因素。初步结果显示，细胞培养在 ECM Matrigel采用使人联想到乳房三维形态 在体内的结构和显示不同的敏感性Fas介导的 凋亡有机氯破坏粘附的水平和关联 与细胞-ECM和细胞-细胞通讯相关的分子。治疗 细胞与p，p '-DDT导致增加的δ-连环蛋白水平相对于 对照组，而Kepone使α-连环蛋白表达降低2倍， 相对于对照组的桥粒。这项研究的假设是， 外源性物质改变细胞粘附的水平、定位和关联 对形成粘着斑、桥粒和粘附物至关重要的分子 连接，其最终影响细胞内信号传导途径（例如PI 3-激酶IAkt激酶），基因表达，细胞周期调控，增殖和 凋亡我们将使用有机氯（p，p '-DDT、o，p'-DDT和甲酮）， 非有机氯异生物质（DMBA，TPA，PhIP）。因此 这项研究的具体目的，采用乳腺上皮细胞培养， Matrigel，是：1）为了确定异生素对分布的影响， 正常细胞-ECM相互作用关键的细胞粘附分子的关联 和局灶性粘连、粘连连接或桥粒。2)来检测异生物质 对PI 3-激酶/Akt信号转导的影响。3)为了确定外源性物质对 细胞周期和细胞周期调节组分（pRB，E2 F，p53，细胞周期蛋白和 细胞周期蛋白依赖性激酶（CDK））。4)为了检测外源性物质是否会改变 Fas介导的凋亡。5)为了评估异生物质浓度是否 与体内组织水平相关，使用 微阵列分析。该提案的结果将提供新颖的 关于乳腺上皮细胞中化学介导效应的信息， 人类增生性乳腺基本细胞变化的精液数据 疾病