KLF8 Regulation of EMT Tumor Metastasis

KLF8对EMT肿瘤转移的调节

基本信息

批准号：
8256614
负责人：
JIHE ZHAO
金额：
$ 10.4万
依托单位：
UNIVERSITY OF CENTRAL FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-01 至 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8256614
关键词：
Acinus organ component Adhesions Biochemical Bioluminescence Breast Cancer Cell Cause of Death Cells E-Cadherin Epithelial Cells Event Fatty acid glycerol esters Focal Adhesion Kinase 1 Genetic Genetic Transcription Goals Human Image Injection of therapeutic agent Integrins Kruppel-like transcription factors Lung MMP2 gene MMP9 gene Mammary gland Matrix Metalloproteinases Mediating Modeling Molecular N-Cadherin Neoplasm Metastasis Nude Mice Oncogenic Pathway interactions Patients Phenotype Pilot Projects Play Proteins RNA Regulation Repression Role Signal Transduction Snails Sorting - Cell Movement Specimen Tail Testing Tumor Cell Invasion Variant Veins Woman Xenograft procedure cancer cell catenin p120ctn protein design epithelial to mesenchymal transition human MMP14 protein in vivo in vivo Model knock-down malignant breast neoplasm mouse model neoplastic cell novel novel therapeutic intervention novel therapeutics overexpression prevent slug tumor tumor progression

项目摘要

Project summary Breast cancer remains one of the leading causes of death in women and most patients die from the tumor metastasis. Our long-term goal is to understand the molecular mechanisms of metastasis so that novel therapeutic strategy can be designed. EMT (epithelial to mesenchymal transition) is a determining step for a cancer cell to progress from a non-invasive to invasive state. Disruption of intercellular adhesion via loss of E- cadherin, the defining event for EMT, is often correlated with gain of the matrix metalloproteinases (MMP) that catalyze proteolytic matrix remodeling important for tumor cell invasion. Such a correlation has been found in invasive breast cancer. However how EMT is regulated during and contributes to breast cancer metastasis is largely unknown. We previously identified Kr¿ppel-like transcription factor 8 (KLF8) as a downstream effector of FAK that plays a crucial role in breast cancer metastasis. Recently, we found that KLF8 is important for oncogenic transformation and overexpressed in invasive human breast cancer, suggesting an uninvestigated role for KLF8 in breast cancer metastasis. Our preliminary studies, using inducible and constitutive expression of KLF8 in MCF-10A cells, suggest that KLF8 is a potent inducer of EMT and invasion by repressing E- cadherin and activating MMP9, MMP2 and MT1-MMP. We also show that KLF8 plays a key role in the loss of E-cadherin and gain of the MMPs in invasive human breast cancer cells and that aberrant KLF8 elevation is tightly correlated with the loss of E-cadherin in patient specimens. Finally, our pilot studies suggest a role for KLF8 in supporting MDA-MB-231 lung metastasis. These results strongly support our hypothesis that KLF8 promotes breast cancer metastasis by initiating EMT and invasion through repression of E-cadherin and activation of the MMPs. We propose three specific aims to test this hypothesis using in semi-in vivo 3D acinus culture and in vivo mouse models and our MCF-10A cells expressing inducible KLF8 and MDA-MB-231 cells expressing inducible KLF8 sh RNA. In Aim 1, we will determine the role of E-cadherin and the MMPs in KLF8- induced EMT and invasion by genetically modifying these proteins. In Aim 2, we will identify the molecular mechanisms by which E-cadherin, the MMPs and intergrin-FAK signaling interplay to regulate KLF8-induced EMT and invasion using genetic, biochemical or pharmacological approaches. In Aim 3, we will determine the role of KLF8 in tumor metastasis using a mouse model of human breast cancer and bioluminescence imaging. We will also evaluate role for other EMT regulators such as Snails, ZEBs and Twist in the regulation of metastasis by KLF8. Completion of this project will shed new lights on mechanisms responsible for breast cancer metastasis which may leads to new therapeutic intervention.

项目概要乳腺癌仍然是女性死亡的主要原因之一，大多数患者死于肿瘤我们的长期目标是了解转移的分子机制，以便发现新的转移。可以设计治疗策略（EMT）（上皮间质转化）。通过 E- 的丧失，癌细胞从非侵袭性状态发展到侵袭性状态。钙粘蛋白是 EMT 的决定性事件，通常与基质金属蛋白酶 (MMP) 的增益相关，催化蛋白水解基质重塑对肿瘤细胞侵袭很重要。然而，EMT 在乳腺癌转移过程中的调节以及如何促进乳腺癌转移尚不清楚。我们之前基本上不知道Kr¿。 ppel 样转录因子 8 (KLF8) 作为下游效应子最近，我们发现 KLF8 对于乳腺癌转移很重要。致癌转化并在侵袭性人类乳腺癌中过度表达，表明未经研究的我们利用诱导型表达和组成型表达进行了初步研究。 KLF8 在 MCF-10A 细胞中的表达，表明 KLF8 通过抑制 E- 是 EMT 和侵袭的有效诱导剂。我们还表明，KLF8 在钙粘蛋白的丧失中发挥着关键作用。 E-钙粘蛋白和侵袭性人乳腺癌细胞中 MMP 的增益以及异常的 KLF8 升高是与患者标本中 E-钙粘蛋白的损失密切相关最后，我们的初步研究表明了其作用。 KLF8 支持 MDA-MB-231 肺转移这些结果有力地支持了我们的假设：KLF8。通过启动 EMT 促进乳腺癌转移，并通过抑制 E-钙粘蛋白和侵袭来促进乳腺癌转移我们提出了三个具体目标来使用半体内 3D 腺泡来检验这一假设。培养和体内小鼠模型以及表达诱导型 KLF8 和 MDA-MB-231 细胞的 MCF-10A 细胞表达诱导型 KLF8 sh RNA 在目标 1 中，我们将确定 E-钙粘蛋白和 MMP 在 KLF8- 中的作用。通过基因修饰这些蛋白质来诱导 EMT 和侵袭。在目标 2 中，我们将鉴定分子。 E-钙粘蛋白、MMP 和整合素-FAK 信号相互作用调节 KLF8 诱导的机制在目标 3 中，我们将使用遗传、生化或药理学方法来确定 EMT 和侵袭。使用人类乳腺癌小鼠模型和生物发光成像研究 KLF8 在肿瘤转移中的作用。我们还将评估其他 EMT 监管机构（例如 Snails、ZEB 和 Twist）在监管中的作用该项目的完成将为乳腺癌的机制提供新的线索。癌症转移可能会导致新的治疗干预。