ZINC DISTRIBUTION IN PROSTATE CELL LINES

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。列出的机构是 中心，不一定是研究者的机构。 我们的研究集中在非癌细胞系（RWPE-1）和三个癌细胞系（DuCaP，VCaP和LnCaP）。 这些前列腺癌细胞系中的两种（DuCaP和VCaP）代表源自侵袭性肿瘤的新型细胞系。 我们已经使用电感耦合质谱法（ICP-MS）定义了这些细胞中的总Zn 2+水平，并证明了癌细胞系显示出总Zn 2+的显著降低以及在用外源性金属处理后不能浓缩Zn 2+。 我们工作的主要重点是开发和使用荧光传感器和活细胞荧光显微镜来成像细胞不同区域中的不稳定Zn 2+。 我们已经发现癌细胞和非癌细胞之间存在显着差异;然而，我们只有一些亚细胞位置的探针。 这项研究将大大加强发展一个全面的图片锌定位在细胞中。 为了理解为什么前列腺癌细胞表现出比健康细胞更少的Zn，我们想要追求使我们能够定义Zn是否存在整体均匀减少的测量，或者Zn是否从特定的亚细胞位置耗尽。 幸运的是，micro-XRF方法作为活细胞成像的补充，有可能产生重大影响。我们的目标是绘制癌性与非癌性前列腺细胞中的锌水平和分布，以确定疾病进展中发生的细胞和亚细胞变化。 这是确定锌在正常前列腺中的作用以及阐明锌缺乏如何与癌症发病相关的关键第一步。 基于同步辐射的X射线荧光微探针测量非常适合于在固定细胞中以适当的长度尺度定量绘制锌的形貌和异质性。 此外，这将补充我们的活细胞研究。

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. Our study focuses on a non-cancerous cell line (RWPE-1) and three cancerous lines (DuCaP, VCaP, and LnCaP). Two of these prostate cancer lines (DuCaP and VCaP) represent novel cell lines derived from aggressive tumors. We have defined the total Zn2+ levels in these cells using Inductively Coupled Mass Spectrometry (ICP-MS) and demonstrate that the cancer cell lines display a marked reduction in total Zn2+ as well as an inability to concentrate Zn2+ upon treatment with exogenous metal. The main focus of our work has been to develop and use fluorescent sensors and live-cell fluorescence microscopy to image labile Zn2+ in different regions of the cell. We have discovered that there are significant differences between the cancerous and non-cancerous cells; however, we only have probes for some subcellular locations. This study would be greatly strengthened by developing a comprehensive picture of Zn localization in cells. To understand why prostate cancer cells exhibit less Zn than healthy cells, we would like to pursue measurements that enable us to define whether there is an overall homogenous reduction in Zn, or whether Zn is depleted from specific sub-cellular locations. Fortunately, micro-XRF methods have the potential for significant impact as a complement to live-cell imaging. Our goal is to map zinc levels and distribution in cancerous vs. non-cancerous prostate cells in order to define the cellular and sub-cellular changes that occur in disease progression. This is a critical first step in defining the role zinc plays in normal prostate and elucidating how zinc depletion correlates with the onset of cancer. Synchrotron-based X-ray fluorescence microprobe measurements are ideally suited to quantitatively map zinc topography & heterogeneity on an appropriate length scale in fixed cells. Additionally, this would compliment our live cell studies.