TARGETS OF AMINO ACID RESTRICTION IN PROSTATE CANCER

前列腺癌中氨基酸限制的目标

• 批准号: 7393213
• 负责人: GARY G MEADOWS
• 金额: $ 25.68万
• 依托单位: WASHINGTON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-06-02 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7393213
• 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; Invasive; 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; Prostate; 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; migration; research study; rho; tool

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） 本申请的目的是鉴定特定氨基酸依赖性调节人雄激素非依赖性前列腺癌细胞的活力和侵袭性的分子靶标。我们推测，特定的氨基酸调节的入侵是依赖于抑制FAK及其结合伙伴。我们进一步假设，特定的氨基酸调控的细胞凋亡诱导是由于MEKJERK存活途径和Akt途径之间的串扰的调节和/或干扰，导致线粒体完整性的丧失，从而激活效应半胱天冬酶。具体目标是：1）鉴定FAK/Cas/Crk或Rho/Ras途径是否是整合素介导的附着/侵袭中的特异性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生存途径之间的串扰。3)确定Bcl-2蛋白家族的BH 123和/或BH 3蛋白对线粒体外膜透化（MOMP）、细胞色素c的线粒体释放和凋亡诱导因子（AIF）以及随后的半胱天冬酶活化的作用，和4）确定饮食酪氨酸和苯丙氨酸限制和甲硫氨酸限制是否将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包括细胞附着、迁移/侵袭和创伤测定以及免疫沉淀和Western印迹分析。将用共聚焦免疫荧光显微镜检查各种信号分子的细胞位置。基因转染实验将用于确定某些细胞信号分子的作用。将通过高压液相色谱法测定细胞内氨基酸，并通过流式细胞术测量细胞凋亡。 拟议研究的一个主要好处是，它将扩展知识到前列腺癌细胞特异性凋亡研究的新途径，并增强对酪氨酸/苯丙氨酸和甲硫氨酸限制的抗癌活性机制的理解。这是特别重要的研究，因为仍然没有令人满意的药物用于治疗雄激素非依赖性转移性人前列腺癌。这项研究可以作为未来开发更特异性的抗转移，抗侵袭，基于前列腺癌的治疗方法的基础。