Oestrogen-related receptor alpha inverse agonist XCT-790 arrests A549 lung cancer cell population growth by inducing mitochondrial reactive oxygen species production
Oestrogen-related receptor alpha inverse agonist XCT-790 arrests A549 lung cancer cell population growth by inducing mitochondrial reactive oxygen species production
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DOI：
10.1111/j.1365-2184.2009.00659.x
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发表时间：
2010-04-01
影响因子：
8.5
通讯作者：
Wong, C.
中科院分区：
文献类型：
--
作者：
Wang, J.;Wang, Y.;Wong, C.
目的：虽然雌激素相关受体 α (ERR α) 主要被认为调节能量稳态，但它也可作为癌症的预后标志物。本研究的目的是调查 ERR α 活性与细胞群生长之间的联系。 材料和方法：XCT-790 是一种 ERRa 特异性反向激动剂，用于抑制人非小细胞肺癌细胞 (NSCLC) A549 中的 ERRa 活性。使用定量实时PCR和蛋白质印迹分析检测基因表达。通过分别用 Mitotracker green、JC-1 和 CM-H(2)DCFDA 染料染色来测量线粒体质量、膜电位和活性氧 (ROS) 产生。通过测量柠檬酸合酶和琥珀酸脱氢酶的活性来分析三羧酸（TCA）循环的进展速率。使用流式细胞术进行细胞周期分析。结果：我们发现 XCT-790 处理减少了线粒体质量，但通过增加 TCA 循环速率、提高线粒体膜电位 (Delta Psi(m)) 和下调超氧化物歧化酶的表达来增强线粒体 ROS 产生。进一步证明XCT-790诱导的ROS可调节p53和Rb信号通路并抑制细胞复制。结论：ERR α通过调节线粒体质量和功能影响细胞周期机制。这一重要途径的失调会导致线粒体 ROS 产生增加，进而调节肿瘤抑制因子的活性，导致细胞周期停滞。
Objective:Although oestrogen-related receptor alpha (ERR alpha) is primarily thought to regulate energy homeostasis, it also serves as a prognostic marker for cancer. The aim of this study was to investigate any connection between ERR alpha activity and cell population growth.Materials and methods:XCT-790, an ERRa specific inverse agonist, was employed to suppress ERRa activity in human non-small cell lung cancer cells (NSCLC) A549. Gene expressions were detected using quantitative real-time PCR and Western blot analysis. Mitochondrial mass, membrane potential and reactive oxygen species (ROS) production were measured by staining with Mitotracker green, JC-1 and CM-H(2)DCFDA dyes respectively. Rate of progression through the tricarboxylic acid (TCA) cycle was analysed by measuring activities of citrate synthase and succinate dehydrogenase. Cell cycle analysis was performed by using flow cytometry.Results:We found that XCT-790 treatment reduced mitochondrial mass but enhanced mitochondrial ROS production by increasing rate through the TCA cycle, elevating mitochondrial membrane potential (Delta Psi(m)) and down-regulating expression of superoxide dismutase. It was further demonstrated that XCT-790-induced ROS modulated p53 and Rb signalling pathways and suppressed cell replication.Conclusions:ERR alpha affects cell cycle mechanisms through modulating mitochondrial mass and function. Dysregulation of this essential pathway leads to elevation in mitochondrial ROS production, which in turn modulates activities of tumour suppressors, resulting in cell cycle arrest.