NICKEL AND HOMOCYSTEINE RESPONSIVE GENE CHARACTERIZATION

P43是一种新发现的功能未知的基因，由 培养的人脐静脉内皮细胞同型半胱氨酸 此外，镍还存在于包括人脐静脉内皮细胞在内的各种人类细胞中。 从长远来看，对该基因功能的研究可能会阐明 镍的重要性或毒性所涉及的机制(例如 致癌)。此外，血浆同型半胱氨酸水平高是 在人类中与动脉硬化有关，在大鼠中这种疾病 在服用镍后被观察到。归纳法的研究 镍和同型半胱氨酸对P43的影响可能会增加我们对 这种疾病的发病机制。短期目标是理解 P43基因产物的功能及镍如何诱导该基因 在培养的细胞中。具体目标是：(1)确定效果 P43在细胞功能上的低表达或高表达 反义或强启动子改变p43和p43转录速率 通过Northern印迹分析、免疫细胞化学等方法监测诱导 研究P43蛋白在细胞内的分布，并进行高效液相和薄层色谱分析 检测细胞内同型半胱氨酸代谢；(2)研究 镍或同型半胱氨酸对细胞同型半胱氨酸代谢的影响 正常细胞和转化细胞及镍对细胞的生化影响 同型半胱氨酸代谢通过蛋氨酸的酶分析。

P43 is a newly discovered gene of unknown function that is induced by homocysteine in cultured human umbilical vein endothelial cells (HUVEC) and also by nickel in a variety of human cell types including HUVEC. In the long term, research on this gene's function may elucidate mechanisms involved in nickel's essentiality or toxicity (e.g. carcinogenesis). In addition, high plasma homocysteine levels are associated with arteriosclerosis in humans and this disease in rats has been observed following nickel administration. Studies of the induction of P43 by nickel and homocysteine may increase our understanding of the pathogenesis of this disorder. The short term goals are to understand the functions of the P43 gene product and how nickel induces the gene in cultured cells. The specific aims are to (1) determine the effects of under or overexpression of P43 on cellular functions by using antisense or a strong promoter to alter transcription rates of P43 and monitoring induction by Northern blot analysis, immunocytochemistry to study cellular distribution of the P43 protein, and HPLC and TLC to examine intracellular homocysteine metabolism; (2) study the effects of nickel or homocysteine doses on cellular homocysteine metabolism in normal and transformed cells and the biochemical effects of nickel upon homocysteine metabolism via enzymatic assays of methionine.