METHOTREXATE UPTAKE AND METABOLISM IN HEPATIC CELLS

这项拟议的研究旨在阐明维生素C的转运和代谢。 肝组织中的甲氨蝶呤和叶酸辅酶。单层文化 大鼠肝细胞系统和大鼠肝癌(H35)细胞系将用于 检查传输过程。将对以下各项进行比较评估： 将对正常细胞、转化细胞和正常细胞进行检查 确定允许这些像元充当的存储位置的属性 叶酸。已培育出H35细胞的耐运输系，并 将该细胞的特性与正常H35细胞进行比较。两者都有 肝细胞特别是H35细胞催化甲氨蝶呤的快速转化 到聚谷氨酸衍生品。将对该反应的性质进行研究 用培养中的细胞和无细胞提取液来了解酶和 对反应的调节。我们还将评估 多聚谷氨酸对甲氨蝶呤在H35细胞中的化疗作用 结合到大分子载体上的甲氨蝶呤将被检查以确定 它们对正常和耐药的H35细胞的比较有效性。

The proposed study is designed to elucidate the transport and metabolism of methotrexate and the folate coenzymes in hepatic tissue. A monolayer culture system of rat hepatocytes and a rat hepatoma (H35) cell line will be used to examine the transport processes. A comparative assessment will be made between the mormal and transformed cells and the normal cells will be examined to determine the properties that allow these cells to act as the storage site for folates. A transport resistant line of the H35 cells has been developed and the properites of the cells will be compared with the normal H35 cells. Both hepatocytes and especially H35 cells catalyze a rapid conversion of methotrexate to polyglutamate derivatives. The properties of this reaction will be studied with cells in culture and cell free extracts to understand the enzymology and regulation of the reaction. We will also assess the contribution that the polyglutamates make to the chemotherapeutic action of methotrexate in H35 cells. Methotrexate bound to macromolecular carriers will be examined to determine their comparative effectiveness against normal and resistant H35 cells.