Characterization of zinc dysregulation in normal and cancerous prostate cells.

说明(申请人提供)：锌是一种重要的微量元素，存在于人体各处，在结构和催化活动中发挥作用。它是人体内第二丰富的过渡金属。前列腺中的锌离子浓度最高。虽然正常的前列腺细胞有异常高的锌+2水平，但前列腺癌患者的锌+2水平明显下降。目前尚不清楚锌离子减少是否是前列腺癌的原因或后果，但了解非癌细胞和癌细胞中锌离子分布的差异是很重要的。这项研究的总体目标是确定非癌和癌前列腺细胞中锌离子池的差异，并将这些变化与调节锌离子稳态的蛋白质相关联。因为在前列腺癌中，锌离子显著减少，所以我们想要阐明细胞中锌离子在哪里减少。细胞有能力在分泌囊泡中储存不稳定的锌离子，并能够对某些细胞信号，如压力或细胞信号做出反应，动员这种金属。本研究旨在揭示非癌细胞和癌细胞中锌离子在亚细胞水平上的分布差异。其目的是确定锌离子的生理作用，以及确定锌离子还原是否在前列腺癌的病理过程中起作用。这一建议有两个具体目标(1)表征负责控制锌离子稳态的蛋白质的表达水平和/或定位的变化；(2)使用针对细胞不同隔室的遗传编码传感器来阐明非癌症和癌症细胞系中的锌+2池。这些活细胞研究将得到固定细胞上的X射线荧光显微镜的补充，以绘制亚细胞部分的总锌离子和电感耦合等离子体质谱。 公共卫生相关性：对非癌和癌前列腺细胞中的锌+2离子进行活细胞成像将极大地增强我们对正常前列腺细胞癌变时这种金属是如何失调的认识。这项工作中开发的工具可以针对细胞的不同隔室，以获得单个细胞器中锌+2的定量水平。获得不同隔室中锌离子的定量水平将开启新的研究领域，以探索哪些蛋白质参与了锌离子结合和细胞信号。

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.