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Protein Kinase C Substrates in Human Breast Cancer

人类乳腺癌中的蛋白激酶 C 底物

基本信息

批准号：
8495571
负责人：
Susan A. Rotenberg
金额：
$ 46.5万
依托单位：
QUEENS COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-02-01 至 2016-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8495571
关键词：
3-Dimensional Affect Affinity Animals Behavior Binding Binding Proteins Biological Assay Biological Markers Bioluminescence Breast Breast Cancer Cell Cells Complex Consensus Detection Diagnostic Disease EGF gene Enzymes Epidermal Growth Factor Evaluation Event Fatty acid glycerol esters Foundations Gap Junctions Growth Human IL8 gene Impact evaluation Implant In Vitro Intervention Investigation Laboratories Lactones Lead Life Link Luciferases Mammary Neoplasms Mediating Methods Microtubule Stabilization Microtubules Modeling Mutate Neoplasm Metastasis Nude Mice Outcome Pathway interactions Pharmaceutical Preparations Phase Phenotype Phosphorylation Phosphorylation Site Phosphotransferases Protein Kinase C Proteins Reading Research Resistance Role Signal Pathway Signal Transduction Testing Tubulin Tumor Cell Line Work cancer cell cell cortex cell motility cell transformation cellular imaging chemical genetics chemotherapy comparative design in vitro Assay in vivo malignant breast neoplasm monomer mouse model mutant neoplastic cell novel novel strategies polarized cell public health relevance response tumor tumor growth

项目摘要

DESCRIPTION (provided by applicant): Protein kinase C (PKC) is an upstream component of the EGF-stimulated (erbB2) and IL-8-dependent signaling pathways whose activity has been linked to motile behavior of cultured human breast cells and to the acquisition of metastatic breast cancer. Therefore, in addition to PKC itself, identification of PKC substrate proteins would offer potential targets for developing anti-metastasis chemotherapies. This laboratory recently discovered that ¿-tubulin and Cdc42 effector protein-4 (CEP4) are substrates of PKC in human breast cells. Phosphorylation of ¿-tubulin at Ser-165 modulates microtubule dynamics such that the growth phase is prolonged, which correlates with cell movement. The significance of ¿-tubulin phosphorylation to microtubule dynamics and cell movement will be investigated by testing phosphorylation site mutants of ¿6-tubulin both in non-transformed MCF-10A cells and in highly metastatic human cancer cells (Specific Aim 1). The mechanistic significance of CEP4 phosphorylation will be explored with phosphorylation site-specific mutants in phenotypes of human breast cells (Specific Aim 2). Since CEP4 binds Cdc42 with high affinity, special emphasis will be on determining how CEP4 phosphorylation affects this binding interaction and consequently how Cdc42 signaling is affected. In addition, a role for IQGAP will be explored in the PKC signaling pathway. IQGAP, a known PKC substrate that requires activated Cdc42 for signaling, is also a required component for MT capture at the cell cortex. Re-localization of IQGAP to lamellapodia occurs in response to PKC activation and be used as a read-out for pro-motility signals arising from CEP4 phosphorylation and activated Cdc42. Thus, IQGAP may provide a nexus at which elongating microtubules and the CEP4-Cdc42 complex converge to promote motility. The significance of the three PKC substrates to tumor growth and metastasis will be explored with phosphorylation site-specific mutants expressed by MDA-MB-231 cells in three dimensional culture assays in vitro (Specific Aim 3). The 3-D assays will guide the selection of which substrate to analyze in an orthotopic nude mouse model. For the in vivo studies, the impact of a phosphorylation-resistant PKC substrate will be evaluated in transfectants of luciferase-MDA-MB-231 cells implanted in the fat pads of nude mice. Tumor formation, growth, and metastasis will be tracked by whole animal bioluminescence (Specific Aim 3). Overall, the results of these studies will establish a coherent mechanistic model that incorporates three PKC substrates (¿-tubulin, CEP4, and IQGAP), and provides a foundation for further investigations of the motility pathway. The anticipated findings are likely to inform strategies for design of novel chemotherapeutics for breast cancer, and to provide new bio-markers for predicting metastatic potential.

描述（由申请人提供）：蛋白激酶C （PKC）是egf刺激（erbB2）和il -8依赖的信号通路的上游组分，其活性与培养的人乳腺细胞的运动行为和转移性乳腺癌的获得有关。因此，除了PKC本身，鉴定PKC底物蛋白将

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Phosphorylation state of Ser165 in α-tubulin is a toggle switch that controls proliferating human breast tumors.

α-微管蛋白中 Ser165 的磷酸化状态是控制增殖的人类乳腺肿瘤的切换开关。

DOI：
10.1016/j.cellsig.2018.08.021
发表时间：
2018
期刊：
Cellular signalling
影响因子：
4.8
作者：
Markovsky,Ela;deStanchina,Elisa;Itzkowitz,Aryeh;Haimovitz-Friedman,Adriana;Rotenberg,SusanA
通讯作者：
Rotenberg,SusanA

PhosphoMARCKS drives motility of mouse melanoma cells.