Journal of Inflammation Research (Oct 2023)
Analysis of Human microRNA Expression Profiling During Diquat-Induced Renal Proximal Tubular Epithelial Cell Injury
Abstract
Yang Chen,1 Hui-Yi Li,1 Jian-Shu Liu,1 Dao-long Jiang,1 Hao-nan Zheng,2 Xue-Song Dong1 1Department of Emergency, The First Hospital of China Medical University, Shenyang, 110001, People’s Republic of China; 2No.105 Phase, The First Clinical College of China Medical University, Shenyang, 110001, People’s Republic of ChinaCorrespondence: Xue-Song Dong, Department of Emergency, The First Hospital of China Medical University, No. 155 of Nanjing North Street, Heping District, Shenyang, 110001, People’s Republic of China, Tel +86 024-26111099 ; +86 024-83282011, Email [email protected]: We established a diquat-induced human kidney-2 cells (HK-2 cells) apoptosis model in this study to identify differentially expressed microRNAs (miRNAs) and signaling pathways involved in diquat poisoning via gene sequencing and bioinformatics analysis and explored the related therapeutic benefits.Methods: The effects of diquat on the viability and apoptosis of HK-2 cells were explored using the CCK-8 and Annexin V-FITC/PI double staining methods. Total RNAs were extracted using the TRizol method and detected by Illumina HiSeq 2500. Bioinformatics analysis was performed to explore differentially expressed (DE) miRNAs, their enriched biological processes, pathways, and potential target genes. The RT-qPCR method was used to verify the reliability of the results.Results: Diquat led to HK-2 cell injury and apoptosis played an important role, hence an HK-2 cell apoptosis model in diquat poisoning was established. Thirty-six DE miRNAs were screened in diquat-treated HK-2 cells. The enriched biological process terms were mainly cell growth, regulation of apoptotic signaling pathway, extrinsic apoptotic signaling pathway, and Ras protein signal transduction. The enriched cellular components were mainly cell-cell junction, cell-substrate junction, ubiquitin ligase complex, and protein kinase complex. The enriched molecular functions were mainly Ras GTPase binding, ubiquitin-like protein transferase activity, DNA-binding transcription factor binding, ubiquitin-protein transferase activity, nucleoside-triphosphatase regulator activity, transcription coactivator activity, and ubiquitin-like protein ligase binding. Signaling pathways such as MAPK, FoxO, Ras, PIK3-Akt, and Wnt were also enriched.Conclusion: These findings aid in understanding the mechanisms of diquat poisoning and the related pathways, where DE miRNAs serve as targets for gene therapy.Keywords: apoptosis, biological processes, diquat, HK-2 cells, microRNA
