Leucine-rich repeats and WD repeat domain-containing protein 1 (LRWD1) is implicated in the regulation of signal transduction, transcription, RNA processing and tumor development. However, LRWD1 transcriptional regulation is not fully understood. This study aimed to investigate the relationship between LRWD1 expression and reactive oxygen species (ROS) level in human embryonal carcinoma cell line, NT2/D1 cells, which will help in understanding the transcriptional regulatory role of ROS in cells. Results showed that the exposure of NT2/D1 cells to various concentrations of hydrogen peroxide (H₂O₂) and the nitric oxide (NO) donor sodium nitroprusside (SNP) caused a significant increase in the mRNA and protein expression of LRWD1. In addition, LRWD1 promoter luciferase reporter assay, and Chromatin Immunoprecipitation assay (CHIP assay) showed that nuclear factor erythroid-2-related factor (Nrf2) was involved in the regulation of LRWD1 expression in response to oxidative stress. The involvement of Nrf2 was confirmed by shRNA-mediated knockdown of Nrf2 in NT2/D1 cells, which caused a significant decrease in LRWD1 expression in response to oxidative stress. Similarly, LRWD1 knockdown resulted in the accumulation of H₂O₂ and superoxide anion radical (O2-). Blocking ROS production by N-acetyl cysteine (NAC) protected NT2/D1 shLRWD1cells from H₂O₂-induced cell death. Collectively, oxidative stress increased LRWD1 expression through a Nrf2-dependent mechanism, which plays an important role in cellular adaptation to oxidative stress. These results highlight an evidence, on the molecular level, about LRWD1 transcriptional regulation under oxidative stress.

富含亮氨酸的重复序列和含WD重复域的蛋白1（LRWD1）参与信号转导，转录，RNA加工和肿瘤发展的调控。但是，LRWD1转录调控尚未完全了解。这项研究旨在调查人类胚胎癌细胞系NT2 / D1细胞中LRWD1表达与活性氧（ROS）水平之间的关系，这将有助于理解ROS在细胞中的转录调控作用。结果显示NT2 / D1细胞暴露于各种浓度的过氧化氢（H ₂ O ₂）和一氧化氮（NO）供体硝普钠（SNP）导致LRWD1的mRNA和蛋白质表达显着增加。此外，LRWD1启动子荧光素酶报告基因测定法和染色质免疫沉淀测定法（CHIP测定法）表明，核因子红系-2相关因子（Nrf2）参与了对氧化应激反应的LRWD1表达的调节。shRNA介导的NT2 / D1细胞中Nrf2的敲低证实了Nrf2的参与，这导致LRWD1表达显着下降，以响应氧化应激。同样，LRWD1敲低导致H ₂ O ₂和超氧阴离子自由基（O2-）的积累。阻止N-乙酰半胱氨酸（NAC）保护的ROS产生H ₂ O的NT2 / D1 shLRWD1细胞_2-诱导的细胞死亡。集体，氧化应激通过依赖Nrf2的机制增加了LRWD1的表达，这在细胞对氧化应激的适应中起着重要的作用。这些结果在分子水平上突出了关于氧化应激下LRWD1转录调控的证据。