DIFFERENTIAL CELL SENSITIVITY TO CADMIUM AND CADMIUM-SEQUESTERING MOLECULES

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。子项目及 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。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 本提案的总体目标是了解细胞内引起和防止镉毒性的分子机制。 我们建议使用对镉表现出不同敏感性的细胞系开发一种体外模型，1）研究细胞内参与镉抗性和毒性的分子机制，2）测试新合成的镉螯合分子的细胞毒性。我们已经确定了两个细胞系，具有不同的敏感性镉暴露;人骨肉瘤细胞系（Saos-2）和虹鳟鱼鳃细胞系（RT鳃-W1）。我们将研究镉诱导的细胞死亡的坏死和凋亡，并证明这些细胞系可以作为一个实验模型。与Peter克雷格博士合作，体外模型将用于筛选新合成的镉螯合分子的细胞毒性。Sheryl Hawkes博士将通过进行蛋白质组分析来量化镉和镉螯合分子在两种细胞系中的作用。将研究个别感兴趣的蛋白质，这些蛋白质在镉或镉螯合剂作用下表现出翻译后修饰或上调或下调的变化。 具体测试的目的包括：1）我们将测试RT鳃W1细胞比Saos-2细胞更能抵抗镉诱导的细胞死亡的假设; 2）我们将确定镉螯合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的蛋白表达的差异;我们将确定镉螯合分子是否改变这两种细胞系中的蛋白质表达。

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.