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Transcriptional & cell cycle control by dietary indoles

转录的

基本信息

批准号：
7233983
负责人：
GARY L FIRESTONE
金额：
$ 25.8万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-07-30 至 2009-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7233983
关键词：
Antibodies Breast Cancer Cell Broccoli - dietary Brussels sprouts - dietary CDK2 gene CDKN1A gene Cabbage - dietary Cell Cycle Cell Cycle Arrest Cell Cycle Progression Cell Cycle Proteins Cell Cycle Regulation Cells Class Complementary DNA Complex Condition Cytoplasm DNA Dietary Indole Dimerization Disruption Estrogens Event Gel Gene Expression Genes Genetic Transcription Human In Vitro Indium Indole-3-Carbinol Indoles Malignant Neoplasms Maps Mediating Mitotic Cell Cycle Molecular Mutate Nuclear Nude Mice Nutrient Pathway interactions Phosphotransferases Plasmids Property Proteins Reagent Regulation Reporter Response Elements Role Signal Pathway Signal Transduction Sp1 Transcription Factor Stream Tamoxifen Testing Text Therapeutic Transcriptional Regulation Transfection Tumor-Derived Vegetables base cancer cell cdc Genes cyclin-dependent kinase 6 diindolylmethane human CDK2 protein in vivo indole inhibitor/antagonist novel oncoprotein p21 promoter reproductive size transcription factor

项目摘要

DESCRIPTION (provided by applicant): Indole-3-carbinol (I3C), a naturally occurring compound in vegetables such as cabbage, broccoli and Brussel's sprouts, and its dimerization product 3-3'-diindolylmethane (DIM) are promising chemotherapeutic and chemopreventative agents for human reproductive cancers. We have demonstrated that the direct exposure of human breast cancer cells to I3C or DIM induces a stringent G1 cell cycle arrest through a multistep signaling cascade that controls expression, activity and cellular utilization of key cell cycle components. I3C, but not DIM, down-regulates cyclin dependent kinase-6 (CDK6) expression and promoter activity, whereas, DIM selectively and rapidly stimulates expression and promoter activity of p21Waf1/Cip1, a critical inhibitor of certain CDKs. The indole regulation of cell cycle gene expression result from specific changes in Sp1 transcription factor-promoter interactions. We have recently discovered that I3C, but not DIM, alters the size, composition and subcellular localization (nuclear to cytoplasm) of the CDK2 protein complex, and a novel 85 kDa protein was uncovered that associates with the CDK2 protein complex in an I3C dependent manner. Thus, the transcriptional control of CDK6 and p21Waf1/Cip1, and the posttanslational regulation of CDK2 protein complex utilization are newly defined down-stream targets of the anti-proliferative indole signaling pathway in human breast cancer cells. Our hypothesis is that I3C and DIM induce a G1 cell cycle arrest of human reproductive cancer cells by activating complementary and distinct cascades that that subsequently control the transcription and posttranslational utilization of key cell cycle components. The indole regulated transcription factors, in addition to Spl, that target the CDK6 and p21Waf1/Cip1 promoters will be identified by interactions with indole responsive regions of cell cycle gene promoters, and functionally tested in a cellular text by manipulation of their expression and/or activity. The novel 85 kDa protein that interacts with the CDK2 protein complex in an I3C dependent manner will be defined and characterized for its role in the I3C disruption of the CDK2 protein complex. The actions of the I3C regulated cell cycle components on cancer cell invasion properties, and in the formation of human breast cancer cell-derived tumors will be assessed using in vitro and in vivo strategies. Our collaborative studies represent the first experimental steps necessary to understand the transcriptional and posttranslational mechanisms by which natural indoles control the cell cycle of human breast cancer cells. This information will be particularly valuable to develop new classes of I3C-based therapeutics for reproductive cancers.

说明（由申请人提供）：吲哚-3-甲醇（I3 C），一种天然存在于蔬菜如卷心菜、花椰菜和甘蓝中的化合物，及其二聚产物3- 3 '-二吲哚基甲烷（DIM）是用于人类生殖系统癌症的有希望的化疗剂和化学预防剂。我们已经证明，人乳腺癌细胞直接暴露于I3 C或DIM诱导严格的G1期细胞周期阻滞通过多步信号级联，控制表达，活性和细胞利用的关键细胞周期成分。I3 C而非DIM下调细胞周期蛋白依赖性激酶6（CDK 6）表达和启动子活性，而DIM选择性地且快速地刺激p21 Waf 1/Cip 1（某些CDK的关键抑制剂）的表达和启动子活性。吲哚对细胞周期基因表达的调控是Sp1转录因子-启动子相互作用的特异性改变的结果。我们最近发现，I3 C，而不是DIM，改变CDK 2蛋白复合物的大小，组成和亚细胞定位（细胞核到细胞质），并发现了一种新的85 kDa的蛋白质，与CDK 2蛋白复合物在I3 C依赖性的方式。因此，CDK 6和p21 Waf 1/Cip 1的转录控制，以及CDK 2蛋白复合物利用的转录后调节是新定义的人乳腺癌细胞中抗增殖吲哚信号通路的下游靶点。我们的假设是，I3 C和DIM诱导G1期细胞周期停滞的人生殖癌细胞通过激活互补的和不同的级联，随后控制转录和翻译后利用的关键细胞周期成分。除了Spl之外，靶向CDK 6和p21 Waf 1/Cip 1启动子的吲哚调节的转录因子将通过与细胞周期基因启动子的吲哚响应区的相互作用来鉴定，并通过操纵它们的表达和/或活性在细胞试验中进行功能测试。将定义以I3 C依赖性方式与CDK 2蛋白复合物相互作用的新型85 kDa蛋白，并表征其在CDK 2蛋白复合物的I3 C破坏中的作用。将使用体外和体内策略评估I3 C调节的细胞周期组分对癌细胞侵袭特性和人乳腺癌细胞衍生肿瘤形成的作用。我们的合作研究代表了理解天然吲哚控制人类乳腺癌细胞细胞周期的转录和翻译后机制所必需的第一个实验步骤。这些信息对于开发新的基于I3 C的生殖系统癌症治疗方法特别有价值。