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Glutathione S-Transferase Functions in Chemoprevention

谷胱甘肽 S-转移酶在化学预防中的作用

基本信息

批准号：
7900820
负责人：
ALAN J TOWNSEND
金额：
$ 8.08万
依托单位：
WAKE FOREST UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-03 至 2010-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7900820
关键词：
4 hydroxynonenal ABCC1 gene Abbreviations Affect Aflatoxin B1 Apoptosis Aromatic Polycyclic Hydrocarbons Benzo(a)pyrene Biological CYP1A1 gene CYP1B1 gene Carcinogens Cell Communication Cell Line Cell model Cells Cessation of life Characteristics Chemoprevention Chemopreventive Agent Chemoprotective Agent Chrysenes Coculture Techniques Complex Mixtures Coupled Cytochrome P-450 CYP1A1 Cytochrome P450 DNA Adducts DNA Repair Detection Development Drug Metabolic Detoxication Effectiveness Enzyme Activation Enzymes Estradiol Excision Exposure to Figs - dietary Fluorescence GSTM1 gene GSTP1 gene Genes Genetic Genetic Polymorphism Glutathione S-Transferase Glycols Goals Growth Hand High Pressure Liquid Chromatography Human In Situ Individual Investigation Isoenzymes Knowledge Malignant Neoplasms Measurement Measures Mediating Metabolic Metabolism Methods Multigene Family Mutagenesis Mutate Nature Necrosis Nitroquinolines Oxides P-Glycoproteins Parents Pathway interactions Phase Play Prevention Principal Investigator Progress Reports Proteins Publishing Pyrenes Relative (related person)Research Research Personnel Risk Role Series Specificity Staging Testing Tissues Toxic effect Toxin Transfection Transgenic Organisms Work adduct base carcinogenesis cell type cytotoxic cytotoxicity enantiomer expression vector genotoxicity insight macromolecule member programs protective effect repaired research study response vector

项目摘要

DESCRIPTION (provided by applicant): Research in this lab and others has provided evidence for an important role for expression of glutathione S-transferases (GSTs) (Phase II detoxification) in normal cellular defenses against reactive electrophilic metabolites of carcinogens, and as part of the biological response induced by chemopreventive agents. We have employed a transgenic cell modeling approach to examine the specific contributions of GSTs expressed individually and in combination with relevant genes, including members of the cytochrome P450 (Phase I activation) and the multidrug resistance protein (Phase III efflux) multigene families. We have found that protection against cytotoxicity and/or genotoxicity (DNA adducts or mutagenesis) can be quite different, depending in some cases primarily on GST characteristics and expression level, or in other cases on the nature of the carcinogen, whether it requires activation, and the P450 activation enzyme co-expressed with the GST. We propose to continue these investigations with our existing single-and dual-transfected V79 cell lines that stably express human GSTP1, GSTM1, or GSTA1, alone and also in combination with human P450-1A1 or-1B1, with addition of -1A2. We will focus on a limited set of polycyclic aromatic hydrocarbon (PAH) substrates for these P450s and GSTs, Benzo[a]Pyrene (and its two 7,8-dihydrodiol enantiomers); the more potent DiBenzo[a,I]Pyrene (and its intermediate 11,12-diol metabolites); and 5-methylchrysene. We will also examine metabolism and toxicities of estradiol, reportedly activated by these CYP isozymes and detoxified by hGSTP1, in these cell lines. In Aim #1 we will examine the unique metabolic interactions between each P450 and GST combination, with both cytotoxicity/apoptosis, DNA adducts, and mutagenicity as endpoints. Aim # 2 will model cell-cell interactions, asking whether binary mixtures of these cell lines show simple additivity, or synergy or antagonism in the presence of PAHs (e.g. via exchange of stable intermediates), and if GST expression is more effective in concert with one or the other P450 in the two different cell lines co-cultured. Aim #3 will determine the mechanisms that underlie the large differences observed in GST protection against the cytotoxic vs. genotoxic effects of certain carcinogens. In Aim #4 we will determine the mechanism for the intriguing observation that expression of GSTs with high efficiency for 4-nitroquinoline oxide or 4-hydroxynonenal conjugation confer paradoxical sensitivity instead of protection. These studies will enhance our knowledge of the factors that govern chemoprotective functions of GSTs.

描述（由申请人提供）：本实验室和其他实验室的研究提供了证据，证明谷胱甘肽 S-转移酶 (GST)（第二阶段解毒）的表达在正常细胞防御致癌物的反应性亲电代谢物中发挥着重要作用，并且是化学预防剂诱导的生物反应的一部分。我们采用转基因细胞建模方法来检查单独表达的 GST 以及与相关基因组合表达的具体贡献，包括细胞色素 P450（第一阶段激活）和多药耐药蛋白（第三阶段流出）多基因家族的成员。我们发现，针对细胞毒性和/或基因毒性（DNA 加合物或诱变）的保护可能有很大不同，在某些情况下主要取决于 GST 特征和表达水平，或者在其他情况下取决于致癌物的性质、是否需要激活以及与 GST 共表达的 P450 激活酶。我们建议继续使用我们现有的单转染和双转染 V79 细胞系继续这些研究，这些细胞系单独稳定表达人 GSTP1、GSTM1 或 GSTA1，也可与人 P450-1A1 或 -1B1 组合，并添加 -1A2。我们将重点关注这些 P450 和 GST 的一组有限的多环芳烃 (PAH) 底物：苯并[a]芘（及其两种 7,8-二氢二醇对映体）；更有效的二苯并[a,I]芘（及其中间体11,12-二醇代谢物）；和5-甲基屈。我们还将检查这些细胞系中雌二醇的代谢和毒性，据报道雌二醇被这些 CYP 同工酶激活并被 hGSTP1 解毒。在目标#1中，我们将检查每种 P450 和 GST 组合之间独特的代谢相互作用，以细胞毒性/细胞凋亡、DNA 加合物和致突变性作为终点。目标#2将模拟细胞与细胞之间的相互作用，询问这些细胞系的二元混合物在PAH存在的情况下是否表现出简单的加和性、协同作用或拮抗作用（例如通过稳定中间体的交换），以及在共培养的两种不同细胞系中，GST表达与一种或另一种P450协同作用是否更有效。目标 #3 将确定 GST 针对某些致癌物的细胞毒性与基因毒性作用的保护作用中观察到的巨大差异背后的机制。在目标 #4 中，我们将确定有趣观察的机制，即对 4-硝基喹啉氧化物或 4-羟基壬烯醛缀合高效表达 GST 会带来矛盾的敏感性而不是保护。这些研究将增强我们对控制 GST 化学保护功能的因素的了解。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Cytotoxicity and mutagenicity of dibenzo[a,l]pyrene and (+/-)-dibenzo[a,l]pyrene-11,12-dihydrodiol in V79MZ cells co-expressing either hCYP1A1 or hCYP1B1 together with human glutathione-S-transferase A1.

在与人谷胱甘肽-S-转移酶 A1 共表达 hCYP1A1 或 hCYP1B1 的 V79MZ 细胞中，二苯并[a,l]芘和 (l-)-二苯并[a,l]芘-11,12-二氢二醇的细胞毒性和致突变性。

DOI：
10.1016/j.mrfmmm.2007.04.004
发表时间：
2007
期刊：
Mutation research
影响因子：
0
作者：
Kushman,MaryE;Kabler,SandraL;Ahmad,Sarfaraz;Doehmer,Johannes;Morrow,CharlesS;Townsend,AlanJ
通讯作者：
Townsend,AlanJ

Modeling the metabolic competency of glutathione S-transferases using genetically modified cell lines.

使用转基因细胞系模拟谷胱甘肽 S-转移酶的代谢能力。