TARGETS OF AMINO ACID RESTRICTION IN PROSTATE CANCER

前列腺癌中氨基酸限制的目标

基本信息

批准号：
7908182
负责人：
GARY G MEADOWS
金额：
$ 13.08万
依托单位：
WASHINGTON STATE UNIVERSITY
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：
2009-08-01 至 2011-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7908182
关键词：
Amino Acids Androgens Apoptosis Binding Biological Assay Caspase Cell Survival Cell-Matrix Junction Dependency Development Flow Cytometry Future Genes Growth High Pressure Liquid Chromatography Human Immunofluorescence Microscopy Immunoprecipitation Induction of Apoptosis Integrins Knowledge Location MEKs Malignant neoplasm of prostate Measures Mediating Methionine Mitochondria Molecular Target Neoplasm Metastasis Outer Mitochondrial Membrane PTK2 gene Pathway interactions Pharmaceutical Preparations Phenylalanine Protein Family Proteins Research Role Signal Transduction Signaling Molecule Techniques Transfection Tyrosine Western Blotting Xenograft procedure androgen independent prostate cancer anticancer activity apoptosis inducing factor base cancer cell cytochrome c human CASP4 protein meetings migration research study rho tool

项目摘要

DESCRIPTION (provided by applicant) The objective of this application is to identify the molecular target(s) by which specific amino acid dependency modulates the viability and invasiveness of human androgen-independent prostate cancer calls. We hypothesize that specific amino acid-regulated invasion is dependent on the inhibition of FAK and its binding partners. We further hypothesize that specific amino acid-regulated induction of apoptosis is due to the modulation and/or interference in cross talk between the MEKJERK survival pathway and the Akt pathway leading to loss of mitochondrial integrity with consequent activation of effector caspases. The specific aims are: 1) Identify if FAK/Cas/Crk or Rho/Ras pathway is the molecular target(s) of specific amino acid restriction in integrin-mediated attachment/invasion. 2) Determine how specific amino acid restriction modulates and/or interferes with the cross talk between the MEK/ERK and Akt survival pathways. 3) Determine the role(s) of BH123 and/or BH3 proteins of the Bcl-2 protein family on mitochondrial outer membrane permeabilization (MOMP), mitochondrial release of cytochrome c, and apoptosis-inducing factor (AIF), and subsequent activation of caspases, and 4) Determine if dietary tyrosine and phenylalanine restriction and methionine restriction will inhibit growth and metastasis of prostate cancer xenografts. Specialized techniques utilized in the application involve cell attachment, migration/invasion, and wounding assays and immunoprecipitation and Western blot analysis. The cellular location of various signaling molecules will be examined with confocal immunofluorescence microscopy. Gene transfection experiments will be used to determine the role of certain cell signaling molecules. Intracellular amino acids will be determined by high pressure liquid chromatography and apoptosis will be measured by flow cytometry. A major benefit from the proposed research proposed is that it will expand knowledge into newer pathways of apoptosis research specific for prostate cancer cells as well as enhance understanding of the mechanisms underlying the anticancer activity of tyrosine/phenylalanine and methionine restriction. This is especially important research since there still is no satisfactory drug for treatment of androgen-independent, metastatic human prostate cancer. This research could serve as the basis for future development of more specific antimetastatic, anti-invasive, apoptosis-based therapies for human prostate cancer.

描述（由申请人提供）该应用的目的是确定特定氨基酸依赖性调节人类雄激素独立前列腺癌调用的生存力和侵入性的分子靶标。我们假设特定的氨基酸调节的侵袭取决于FAK及其结合伴侣的抑制。我们进一步假设，特定的氨基酸调节的凋亡诱导是由于Mekjerk存活途径与AKT途径之间的串扰和/或干扰引起的，导致线粒体完整性的丧失，导致效应子caspases的激活。具体目的是：1）确定FAK/CAS/CRK或RHO/RAS途径是整联蛋白介导的附着/侵袭中特定氨基酸限制的分子靶标。 2）确定特定的氨基酸限制如何调节和/或干扰MEK/ERK和AKT存活途径之间的串扰。 3) Determine the role(s) of BH123 and/or BH3 proteins of the Bcl-2 protein family on mitochondrial outer membrane permeabilization (MOMP), mitochondrial release of cytochrome c, and apoptosis-inducing factor (AIF), and subsequent activation of caspases, and 4) Determine if dietary tyrosine and phenylalanine restriction and methionine restriction will抑制前列腺癌异种移植物的生长和转移。应用中使用的专门技术涉及细胞附着，迁移/侵袭以及伤害测定以及免疫沉淀和蛋白质印迹分析。各种信号分子的细胞位置将使用共聚焦免疫荧光显微镜检查。基因转染实验将用于确定某些细胞信号分子的作用。细胞内氨基酸将由高压液相色谱确定，凋亡将通过流式细胞仪测量。拟议的研究提出的一个主要好处是，它将将知识扩展为针对前列腺癌细胞的凋亡研究的新途径，并增强对酪氨酸/苯丙氨酸和蛋氨酸限制抗癌活性的机制的理解。这是特别重要的研究，因为仍然没有令人满意的药物来治疗雄激素独立的转移性人类前列腺癌。这项研究可以作为对人类前列腺癌的基于更具体的抗转移性，抗侵入性，基于凋亡的疗法的未来发展的基础。