MOLECULAR CHARACTERIZATION OF METHADONE RECEPTORS

美沙酮受体的分子表征

基本信息

批准号：
2008212
负责人：
RHODA MANECKJEE
金额：
$ 14.75万
依托单位：
OREGON HEALTH & SCIENCE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1995
资助国家：
美国
起止时间：
1995-01-01 至 1999-12-31
项目状态：
已结题

项目摘要

The long acting synthetic narcotic methadone is a potent inhibitor of the in vivo and in vitro growth of human lung cancer cells and the inhibition appears to be mediated by specific, non-conventional, high affinity methadone binding sites, distinct from the sites present in human brain and normal lung. Based on these results, a clinical trial of methadone therapy for lung cancer patients has been initiated. The goal of the proposed research is (1) to purify, characterize and clone the corresponding genes for methadone receptors present on human lung cancer cells, human brain and normal lung, in order to determine the existence of multiple methadone receptor types in normal and tumor tissues and/or the central (CNS) and peripheral nervous systems (PNS); and (2) to elucidate the molecular mechanisms involved in the growth inhibiting actions of methadone in human lung cancer cells. Purification of the receptors will be carried out using affinity chromatography, and biochemical and pharmacological characteristics of the purified receptors will be determined. Cloning of the methadone receptor expressed in lung cancer cells will be carried out using amino acid sequences of the purified receptor and PCR techniques. The mechanism of action of methadone in human lung cancer cells will be determined by studying (a) its role in the regulation of apoptosis; (b) the signal transduction pathways involved in its apoptotic effect, such as the regulation of protein kinase C activity; and (c) changes in gene expression leading to growth inhibition. These studies should lead to a better understanding of the PNS and CNS effects of this drug and the design of better treatment protocols for its use in the control of cancer growth and pain, as well as in drug abuse programs.

长作用合成麻醉美沙酮是一种有效的抑制剂人体肺癌细胞的体内和体外生长和抑制作用似乎是由特定的，非规定的高亲和力介导的美沙酮结合位点，不同于人类大脑中的位点和正常的肺。基于这些结果，美沙酮的临床试验已经开始针对肺癌患者的治疗。目标的目标拟议的研究是（1）纯化，表征和克隆在人肺癌上存在的美沙酮受体的相应基因细胞，人脑和正常肺，以确定存在正常和肿瘤组织中多种美沙酮受体类型和/或中央（CNS）和周围神经系统（PNS）；（2）到阐明抑制生长的分子机制美沙酮在人肺癌细胞中的作用。纯化受体将使用亲和力色谱法进行，并且纯化受体的生化和药理学特征将确定。在肺中表达的美沙酮受体的克隆癌细胞将使用的氨基酸序列进行纯化的受体和PCR技术。作用机理人肺癌细胞中的美沙酮将通过研究（a）来确定它在调节凋亡中的作用；（b）信号传输涉及其凋亡效应的途径，例如调节蛋白激酶C活性；（c）基因表达的变化领先抑制生长。这些研究应该导致更好了解该药物的PN和CNS效应以及设计更好的治疗方案用于控制癌症生长和痛苦以及药物滥用计划。