Characterization of zinc dysregulation in normal and cancerous prostate cells.

描述（由申请人提供）：锌是一种重要的微量元素，存在于人体各处，具有结构和催化活性。它是人体中第二丰富的过渡金属。在前列腺中发现Zn+2的最高浓度。虽然正常前列腺细胞具有异常高的Zn+2水平，但其在前列腺癌中显著耗尽。目前尚不清楚Zn+2减少是否是前列腺癌的原因或后果，但重要的是要了解非癌细胞和癌细胞之间Zn+2分布的差异。本研究的总体目标是确定非癌性和癌性前列腺细胞之间Zn+2库的差异，并将这些变化与调节Zn 2+稳态的蛋白质相关联。因为Zn+2在前列腺癌中显著减少，我们想阐明细胞中Zn+2减少的位置。细胞具有将不稳定的Zn+2储存在分泌囊泡中的能力，并且能够响应于某些细胞线索（例如应激或细胞信号）而动员这种金属。本研究旨在揭示锌+2在非癌细胞和癌细胞亚细胞水平分布的差异。目的是确定Zn+2的生理作用，以及确定Zn+2减少是否在前列腺癌的病理学中起作用。该提议有两个具体目标：（1）表征负责控制Zn 2+稳态的蛋白质的表达水平和/或定位的变化;（2）使用靶向细胞不同区室的遗传编码传感器阐明非癌细胞系和癌细胞系中的Zn+2库。这些活细胞研究将通过固定细胞的X射线荧光显微镜进行补充，以绘制亚细胞组分的总Zn+2和ICP-MS。 公共卫生相关性：非癌和癌前列腺细胞中Zn+2离子的活细胞成像将极大地增强我们对当正常前列腺细胞变成恶性时这种金属是如何失调的知识。这项工作中开发的工具可以针对细胞的不同隔室，以获得单个细胞器内Zn+2的定量水平。获得定量的Zn+2水平在不同的车厢将打开新的研究领域，以调查哪些蛋白质参与Zn+2结合和细胞信号。

DESCRIPTION (provided by applicant): Zinc is an important trace element that is found throughout the human body and functions in structural and catalytic activities. It is the second most abundant transition metal in the body. The highest concentration of Zn+2 is found in the prostate gland. While normal prostate cells have exceptionally high Zn+2 levels it is markedly depleted in prostate cancer. It is not yet clear if Zn+2 reduction is a cause or consequence of prostate cancer but it is important to understand the differences in Zn+2 distribution between noncancerous and cancer cells. The overall goal of this research is to define differences in Zn+2 pools between noncancerous and cancerous prostate cells and correlate these changes with proteins that regulate Zn2+ homeostasis. Because Zn+2 is markedly reduced in prostate cancer we would like elucidate where in the cell Zn+2 is reduced. Cells have the ability to store labile Zn+2 in secretory vesicles and are able to mobilize this metal in response to certain cellular cues, such as stresses or cellular signals. This study aims to reveal the difference in Zn+2 distributions at the subcellular level in noncancerous compared to cancerous cells. The goal is to define the physiological role of Zn+2, as well as determine whether Zn+2 reduction plays a role in the pathology of prostate cancer. This proposal has two specific aims (1) characterize changes in expression levels and/or localization of proteins responsible for controlling Zn2+ homeostasis; (2) elucidate the Zn+2 pools in noncancerous and cancerous cell lines using genetically encoded sensors targeted to different compartment of the cell. These live cell studies will be complemented by x-ray fluorescence microscopy on fixed cells to map total Zn+2 and ICP-MS on subcellular fractions. PUBLIC HEALTH RELEVANCE: Live cell imaging of Zn+2 ions in noncancerous and cancerous prostate cells will greatly enhance our knowledge on how this metal is dysregulated when normal prostate cells become malignant. The tools developed in this work can be targeted to different compartments of the cells to obtain quantitative levels of Zn+2 within individual organelles. Obtaining quantitative Zn+2 levels in different compartments will open new areas of research to investigate what proteins are involved in Zn+2 binding and cellular signals.