Identification of mitogen-activated protein kinase kinase as a chemoresistant pathway in MCF-7 cells by using gene expression microarray
Identification of mitogen-activated protein kinase kinase as a chemoresistant pathway in MCF-7 cells by using gene expression microarray
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DOI：
10.1067/msy.2002.125389
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发表时间：
2002-08-01
期刊：
影响因子：
3.8
通讯作者：
Burow, ME
中科院分区：
文献类型：
--
作者：
Weldon, CB;Scandurro, AB;Burow, ME
背景。丝裂原活化蛋白激酶（MAPK）级联的组成部分与凋亡调节有关。本研究通过基因表达谱分析，鉴定并暗示裂丝原活化蛋白激酶(MEK5)-大裂丝原活化激酶-1 (bmk1)/细胞外信号相关蛋白激酶（ERK5）通路是参与化疗耐药的新靶点。凋亡敏感（APO+）和凋亡耐药（APO-） MCF-7细胞变体之间的差异基因表达通过芯片检测和逆转录聚合酶链反应（RT-PCR）证实。凋亡/活力报告基因试验用于确定BMK1的显性阴性突变体（BMK1/DN）与凋亡诱导剂（依托泊苷、肿瘤坏死因子- α [TNF]或TNF相关的凋亡诱导配体[TRAIL]），加或不加酚酯（PMA）转染的影响。在通过微阵列分析检测到的1186个基因中，MEK5在APO-细胞中增加了22倍。RT-PCR分析证实MEK5过表达。单独表达BMK1/DN导致细胞死亡与对照组相比呈剂量依赖性增加（P < 0.05）。此外，BMK1/DN增强了MCF-7细胞对治疗诱导的细胞死亡的敏感性（P < 0.05）。PMA部分抑制TRAIL-和tat诱导的细胞死亡的能力被BMK1/DN抑制。然而，只有trail诱导的活性抑制具有统计学意义（P < 0.05）。APO- MCF-7乳腺癌细胞中MEK5的过表达表明，这种MAPK信号蛋白是一种有效的存活分子。MEK5信号的分子抑制可能是癌细胞对化疗方案敏感的一种机制。
Background. Components of the mitogen-activated protein kinase (MAPK) cascade have been implicated in apoptotic regulation. This study used gene expression profiling analysis to identify and implicate mitogen-activated protein kinase kinase (MEK5)-BMK1 (big mitogen-activated kinase-1)/extracellular signal related protein kinase (ERK5) pathway as a novel target involved in chemoresistance.Methods. Differential gene expression between apoptotically sensitive (APO+) and apoptotically resistant (APO-) MCF-7 cell variants was determined by using microarray and confirmed by reverse transcriptase-polymerase chain reaction (RT-PCR). An apoptotic/viability reporter gene assay was used to deter mine the effects of the transfection of a dominant-negative mutant of BMK1 (BMK1/DN) in conjunction with apoptotic-inducing agents (etoposide, tumor necrosis factor-alpha [TNF], or TNF-related apoptosis-inducing ligand [TRAIL]), with or without phorbol ester (PMA).Results. Of the 1186 genes detected through microarray analysis, MEK5 was increased 22-fold in APO- cells. Overexpression of MEK5 was confirmed by using RT-PCR analysis. Expression of BMK1/DN alone resulted in a dose dependent increase in cell death versus control (P < .05). In addition, BMK1/DN enhanced the sensitivity of MCF-7 cells to treatment-induced cell death (P < .05). The ability of PMA to partially suppress TRAIL- and TAT-induced cell death was inhibited by BMK1/DN. However, only TRAIL-induced activity suppression reached statistical significance (P < .05).Conclusions. The overexpression of MEK5 in APO- MCF-7 breast carcinoma cells shows that this MAPK signaling protein represents a potent survival molecule. Molecular inhibition of MEK5 signaling may represent a mechanism for sensitizing cancer cells to chemotherapeutic regimens.