ZINC DISTRIBUTION IN PROSTATE CELL LINES

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 我们的研究集中在一个非癌细胞系(RWPE-1)和三个癌细胞系(DuCaP、VCaP和LNCaP)上。其中两个前列腺癌株(DuCaP和VCaP)代表了来自侵袭性肿瘤的新细胞系。我们已经用电感耦合质谱仪(ICP-MS)确定了这些细胞中的总锌离子水平，并证明了癌细胞株在用外源金属处理后总锌离子显著减少，并且不能浓缩锌离子。我们的主要工作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不稳定的锌离子。我们发现，癌细胞和非癌细胞之间存在着显著的差异，但我们只有一些亚细胞位置的探针。如果能全面了解锌在细胞中的定位，这项研究将得到极大的加强。为了理解为什么前列腺癌细胞比健康细胞表现出更少的锌，我们想要进行一些测量，使我们能够确定是否存在锌的整体同质性减少，或者锌是否从特定的亚细胞位置被耗尽。幸运的是，微型XRF方法有可能作为活细胞成像的补充而产生重大影响。我们的目标是绘制癌症和非癌症前列腺细胞中锌的水平和分布图，以确定疾病进展过程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的第一步。基于同步加速器的X射线荧光微探针测量非常适合于在固定单元中以适当的长度尺度定量绘制锌的形貌和不均一性。此外，这将对我们的活细胞研究起到补充作用。

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.