NICKEL AND HOMOCYSTEINE RESPONSIVE GENE CHARACTERIZATION

P43基因是一个新发现的功能未知的基因，它是由 培养人脐静脉内皮细胞同型半胱氨酸 并且在包括HUVEC在内的多种人类细胞类型中也被镍所影响。 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， 致癌作用）。 此外，高血浆同型半胱氨酸水平 与人类动脉硬化有关，而这种疾病在大鼠中 在镍施用后观察到。 归纳研究 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。 短期目标是了解 P43基因产物的功能以及镍对该基因的诱导作用 在培养的细胞中。具体目标是：（1）确定 P43对细胞功能的影响 反义或强启动子以改变P43的转录速率， 通过北方印迹分析、免疫细胞化学 研究P43蛋白的细胞分布，并进行HPLC和TLC， 检测细胞内同型半胱氨酸代谢;（2）研究 镍或同型半胱氨酸剂量对细胞同型半胱氨酸代谢 正常和转化细胞以及镍对 通过甲硫氨酸的酶测定的同型半胱氨酸代谢。

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.