TARGETS OF AMINO ACID RESTRICTION IN PROSTATE CANCER

前列腺癌中氨基酸限制的目标

• 批准号: 7071292
• 负责人: GARY G MEADOWS
• 金额: $ 26.45万
• 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 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7071292
• 关键词: BCL2 gene /protein; SCID mouse; apoptosis; biological signal transduction; cancer prevention; cell growth regulation; confocal scanning microscopy; cysteine endopeptidases; cytochrome c; dietary aminoacid; dietary restriction; flow cytometry; focal adhesion kinase; high performance liquid chromatography; hormone related neoplasm /cancer; metastasis; methionine; mitochondria; mitogen activated protein kinase; molecular oncology; nutrition aspect of cancer; nutrition related tag; phenylalanine; prostate neoplasms; serine threonine protein kinase; transfection; tyrosine; western blottings

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) 本应用的目的是确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活性和侵袭力的分子靶点(S)。我们假设，特定的氨基酸调节的侵袭依赖于FAK及其结合伙伴的抑制。我们进一步假设，特定氨基酸调控的凋亡诱导是由于MEKJERK生存通路和Akt通路之间的串扰调节和/或干扰导致线粒体完整性丧失，从而激活效应器caspase。其具体目的是：1)确定在整合素介导的黏附/侵袭中，FAK/Cas/Crk或Rho/Ras通路是否是特异性氨基酸限制的分子靶点(S)。2)确定特定的氨基酸限制如何调节和/或干扰MEK/ERK和Akt生存通路之间的串扰。3)确定Bc l-2蛋白家族的BH123和/或BH3蛋白在线粒体外膜通透性、线粒体释放细胞色素c和凋亡诱导因子以及随后激活caspase中的作用(S)；4)确定饮食中酪氨酸和苯丙氨酸限制以及蛋氨酸限制是否会抑制前列腺癌移植瘤的生长和转移。 应用中使用的专门技术包括细胞附着、迁移/入侵、损伤分析、免疫沉淀和Western印迹分析。各种信号分子的细胞位置将用共聚焦免疫荧光显微镜进行检查。基因导入实验将被用来确定某些细胞信号分子的作用。细胞内氨基酸的测定采用高压液相色谱仪，细胞凋亡率的测定采用流式细胞仪。 拟议的研究的一个主要好处是，它将把知识扩展到针对前列腺癌细胞的新的凋亡研究途径，以及加强对酪氨酸/苯丙氨酸和蛋氨酸限制的抗癌活性潜在机制的理解。这项研究尤其重要，因为仍然没有令人满意的药物来治疗雄激素非依赖性、转移性人类前列腺癌。这项研究可以作为未来开发更具特异性的抗转移、抗侵袭、基于细胞凋亡的前列腺癌治疗方法的基础。