V-ATPASE PUMPS IN PROSTATE CANCER: REGULATORY AND FUNCTIONAL STUDIES

前列腺癌中的 V-ATP 酶泵：调节和功能研究

• 批准号: 8167588
• 负责人: Karlett J Parra
• 金额: $ 10.85万
• 依托单位: NEW MEXICO STATE UNIVERSITY LAS CRUCES
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8167588
• 关键词: ATP phosphohydrolase; Acids; Cell model; Cells; Computer Retrieval of Information on Scientific Projects Database; Cytoplasm; Energy Metabolism; Funding; Glucose; Glycolysis; Goals; Grant; Human; Institution; Malignant Neoplasms; Malignant neoplasm of prostate; Measures; Mediating; Metabolism; Neoplasm Metastasis; Normal Cell; Primary Cell Cultures; Prostate; Proton Pump; Pump; Research; Research Personnel; Resources; Small Interfering RNA; Source; Starvation; System; Testing; Tissues; United States National Institutes of Health; Vesicle; cancer cell; extracellular; inhibitor/antagonist; insight; pH Homeostasis; pH gradient; tumor progression; vacuolar H+-ATPase

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. V-ATPases are highly conserved proton pumps important for pH homeostasis. In malignant tumors, V-ATPases sustain an altered pH gradient between the cytoplasm and the lumen of intracellular vesicles; and generate the acid extracellular microenvironment necessary for cancer progression and metastasis. Lack of V-ATPase function blocks cancer progression but the mechanism involved is not known. Our long term goal is to understand how cancer cells regulate V-ATPase pumps and how V-ATPases assist in cancer progression to metastasis. Glucose regulates V-ATPases in normal cells. When glucose is limiting or glycolysis is interrupted the V-ATPases disassemble. Disassembly inactivates the pump and is reversed by glucose readdition. We postulate that glucose also controls V-ATPase assembly and activity in malignant tumors where glycolysis dominates metabolism. To test this hypothesis we will use a cell model of human prostate cancer progression; and human primary cell cultures of normal prostate and prostate cancer tissue. The energy metabolism of the prostate is specialized for fine tuning glycolytic activity which makes the prostate an attractive system for these studies. We will establish parallels between glucose utilization and V-ATPase assembly by measuring reversible disassembly, V-ATPase activity, and cytosolic and lysosomal pH in cells exposed to glucose, glycolytic inhibitors, and starvation (Aim 1). V-ATPase deficient cells, as a result of siRNA-mediated knockdown and treatment with V-ATPase inhibitors, will be used to further discern connections between V-ATPase activity and pH homeostasis (Aim 2). These studies will lend new insight into how V-ATPases aid cancer cells in controlling luminal, cytosolic, and extracellular pH.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 V-ATP酶是高度保守的质子泵，对pH稳态很重要。在恶性肿瘤中，V-ATP酶维持细胞质和细胞内囊泡腔之间改变的pH梯度;并产生癌症进展和转移所必需的酸性细胞外微环境。缺乏V-ATP酶功能会阻止癌症进展，但其机制尚不清楚。我们的长期目标是了解癌细胞如何调节V-ATP酶泵以及V-ATP酶如何帮助癌症进展到转移。 葡萄糖调节正常细胞中的V-ATP酶。当葡萄糖受限或糖酵解中断时，V-ATP酶分解。胰岛素泵失活，并通过葡萄糖再灌注逆转。 我们推测，在糖酵解主导代谢的恶性肿瘤中，葡萄糖也控制V-ATP酶的组装和活性。为了检验这一假设，我们将使用人前列腺癌进展的细胞模型;以及正常前列腺和前列腺癌组织的人原代细胞培养物。前列腺的能量代谢专门用于微调糖酵解活性，这使得前列腺成为这些研究的有吸引力的系统。我们将通过测量暴露于葡萄糖、糖酵解抑制剂和饥饿的细胞中的可逆分解、V-ATP酶活性以及胞质和溶酶体pH来建立葡萄糖利用和V-ATP酶组装之间的平行关系（目的1）。由于siRNA介导的敲低和用V-ATP酶抑制剂处理，V-ATP酶缺陷细胞将用于进一步辨别V-ATP酶活性和pH稳态之间的联系（目的2）。这些研究将为V-ATP酶如何帮助癌细胞控制管腔，胞质和细胞外pH提供新的见解。