Expression of glutathione S-transferase P1-1 in leukemic cells is regulated by inducible AP-1 binding
Expression of glutathione S-transferase P1-1 in leukemic cells is regulated by inducible AP-1 binding
复制标题
DOI：
10.1016/j.canlet.2004.05.004
复制
发表时间：
2004-12-28
期刊：
影响因子：
9.7
通讯作者：
Diederich, M
中科院分区：
文献类型：
--
作者：
Duvoix, A;Schnekenburger, M;Diederich, M
谷胱甘肽s -转移酶（GST）参与细胞对外源性药物、氧化应激的保护以及对化疗化合物（如阿霉素）的耐药性。已知人类胎盘型GSTP1-1水平在许多肿瘤和造血疾病中升高。在这项工作中，我们比较了表达或不表达GSTP1-1的细胞的转录机制。瞬时转染试验表明，不同的gst启动子报告结构产生细胞类型特异性水平的荧光素酶活性。在表达细胞中，转录活性强烈依赖于GSTPI基因-65至-75 bp区域内的AP-1结合元件，如定点突变所示。电泳迁移率转移实验表明，DNA结合活性仅在表达gstp1 -1的细胞中观察到，并且在过氧化氢、TPA、叔丁基对苯二酚和阿霉素刺激后增加。非表达细胞既不存在构成性也不存在诱导性AP-1结合。综上所述，我们的研究结果为白血病细胞通过AP-1结合活性诱导GSTPI基因提供了证据，并有助于更好地理解参与耐药机制的调节基因的分子事件。2004爱思唯尔爱尔兰有限公司版权所有。
Glutathione S-transferases (GST) are involved in cellular protection against xenobiotics, oxidative stress as well as in resistance against chemotherapeutic compounds such as doxorubicin. Levels of human placental type GSTP1-1 are known to be increased in many tumors and hematopoietic diseases. In this work, we compare transcriptional mechanisms in cells that express or not GSTP1-1. Transient transfection assays are used to show that different GST-promoter reporter constructs generate cell-type specific levels of luciferase activity. In expressing cells, transcriptional activity is strongly dependent on AP-1 binding elements within the -65 to -75 bp region of the GSTPI gene as shown by site-directed mutagenesis. Electrophoretic mobility shift assays show that DNA binding activity is exclusively observed in GSTP1-1-expressing cells and is increased after stimulation with hydrogen peroxide, TPA, tert-butylhydroquinone and doxorubicin. Non-expressing cells present neither constitutive nor inducible AP-1 binding. Taken together, our results provide evidence for the induction of the GSTPI gene via AP-1 binding activity in leukemia cells and contribute to a better understanding of the molecular events regulating genes involved in drug resistance mechanisms. (C) 2004 Elsevier Ireland Ltd. All rights reserved.