DIFFERENTIAL CELL SENSITIVITY TO CADMIUM AND CADMIUM-SEQUESTERING MOLECULES

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。 这项建议的总体目标是了解细胞内引起和保护镉毒性的分子机制。我们建议利用对镉表现出不同敏感性的细胞系建立一个体外模型，1)研究细胞内参与镉抗性和毒性的分子机制，2)测试新合成的镉隔离分子的细胞毒性。我们已经鉴定出对镉暴露具有不同敏感性的两种细胞系：人骨肉瘤细胞系(Saos-2)和虹鱼鳃细胞系(RT Gill-W1)。我们将研究镉通过坏死和凋亡诱导的细胞死亡，并证明这些细胞系可以作为实验模型。与彼得·克雷格博士合作，体外模型将用于筛选新合成的镉隔离分子的细胞毒性。谢丽尔·霍克斯博士将通过进行蛋白质组分析来量化镉和镉隔离分子在这两个细胞系中的影响。个别感兴趣的蛋白质将被研究，它们的翻译后修饰或上调或下调对镉或镉隔离剂的响应发生了变化。 测试的具体目标包括：1)我们将检验RT Gill W1细胞比Saos-2细胞对镉诱导的细胞死亡更具抵抗力的假设；2)我们将确定镉隔离分子是否减少镉诱导的细胞死亡；3)我们将比较两种细胞系中镉改变蛋白表达的差异；以及4)我们将确定镉隔离分子是否改变这两种细胞系的蛋白表达。

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. The overall objective of this proposal is to understand the molecular mechanisms within cells that cause and protect against cadmium toxicity. We propose to develop an in vitro model using cell lines that exhibit differential sensitivity to cadmium 1) to study the molecular mechanisms within cells involved in cadmium resistance and toxicity, and 2) to test the cytotoxicity of newly-synthesized cadmium-sequestering molecules. We have identified two cell lines that possess different sensitivity to cadmium exposure; a human osteosarcoma cell line (Saos-2) and a rainbow trout gill cell line (RT gill-W1). We will investigate cadmium induced cell death by necrosis and apoptosis, and demonstrate that these cell lines can be used as an experimental model. In collaboration with Dr. Peter Craig, the in vitro model will be used to screen the cytotoxicity of newly synthesized cadmium-sequestering molecules. Dr. Sheryl Hawkes will quantify the effect of cadmium and cadmium-sequestering molecules in the two cell lines by performing proteome analysis. Individual proteins of interest will be studied that exhibit changes to their post-translational modifications or up- or down-regulation in response to cadmium or cadmium-sequestering agents. The specific aims to be tested include: 1) We will test the hypothesis that RT gill W1 cells are more resistant to cadmium-induced cell death compared to Saos-2 cells; 2) We will determine whether cadmium-sequestering molecules reduce cadium-induced cell death; 3) We will compare the differences in cadmium-altered protein expression in the two cell lines; and 4) We will determine whether cadmium-sequestering molecules alter protein expression in these two cell lines.