Wang Zhi, Liu Yimin, Han Ning, Chen Xuemei, Yu Wei, Zhang Weisen, Zou Fei
School of Public Health and Tropical Medicine, Southern Medical University, No. 1838, North Guangzhou Road, Guangzhou, Guangdong 510515, China.
Brain Res. 2010 Jul 30;1346:14-25. doi: 10.1016/j.brainres.2010.05.059. Epub 2010 May 26.
Oxidative stress and high levels of reactive oxygen species (ROS) are risk factors of auditory cell injury and hearing impairment. MicroRNAs (miRNAs) are critical for the post-transcriptional regulation of gene expression and cell proliferation and survival. However, little is known about the impact of oxidative stress on the expression of miRNAs and their targeted mRNAs in auditory cells. We employed a cell model of oxidative stress by treatment of House Ear Institute-Organ of Corti 1 (HEI-OC1) cells with different concentrations of tert-butyl hydroperoxide (t-BHP) to examine the t-BHP-induced production of ROS and to determine the impact of t-BHP treatment on the relative levels of miRNA and mRNA transcripts in HEI-OC1 cells. We found that treatment with different concentrations of t-BHP promoted the production of ROS, but inhibited the proliferation of HEI-OC1 cells in a dose- and time-dependent manner. Furthermore, treatment with t-BHP induced HEI-OC1 cell apoptosis. Further microarray analyses revealed that treatment with t-BHP increased the transcription of 35 miRNAs, but decreased the expression of 40 miRNAs. In addition, treatment with t-BHP up-regulated the transcription of 2076 mRNAs, but down-regulated the levels of 580 mRNA transcripts. Notably, the up-regulated (or down-regulated) miRNAs were associated with the decreased (or increased) expression of predicted targeted mRNAs. Importantly, these differentially expressed mRNAs belonged to different functional categories, forming a network participating in the oxidative stress-related process in HEI-OC1 cells. Therefore, our findings may provide new insights into understanding the regulation of miRNAs on the oxidative stress-related gene expression and function in auditory cells.
氧化应激和高水平的活性氧(ROS)是听觉细胞损伤和听力障碍的危险因素。微小RNA(miRNA)对基因表达、细胞增殖和存活的转录后调控至关重要。然而,关于氧化应激对听觉细胞中miRNA及其靶向mRNA表达的影响知之甚少。我们通过用不同浓度的叔丁基过氧化氢(t-BHP)处理House Ear Institute-Organ of Corti 1(HEI-OC1)细胞来建立氧化应激细胞模型,以检测t-BHP诱导的ROS产生,并确定t-BHP处理对HEI-OC1细胞中miRNA和mRNA转录本相对水平的影响。我们发现,用不同浓度的t-BHP处理可促进ROS的产生,但以剂量和时间依赖性方式抑制HEI-OC1细胞的增殖。此外,t-BHP处理可诱导HEI-OC1细胞凋亡。进一步的微阵列分析显示,t-BHP处理可增加35种miRNA的转录,但降低40种miRNA的表达。此外,t-BHP处理上调了2076种mRNA的转录,但下调了580种mRNA转录本的水平。值得注意的是,上调(或下调)的miRNA与预测靶向mRNA的表达降低(或增加)相关。重要的是,这些差异表达的mRNA属于不同的功能类别,形成了一个参与HEI-OC1细胞氧化应激相关过程的网络。因此,我们的发现可能为理解miRNA对听觉细胞中氧化应激相关基因表达和功能的调控提供新的见解。