PMID- 29653565 OWN - NLM STAT- MEDLINE DCOM- 20180430 LR - 20240314 IS - 1471-2415 (Electronic) IS - 1471-2415 (Linking) VI - 18 IP - 1 DP - 2018 Apr 13 TI - Identification of H(2)O(2) induced oxidative stress associated microRNAs in HLE-B3 cells and their clinical relevance to the progression of age-related nuclear cataract. PG - 93 LID - 10.1186/s12886-018-0766-6 [doi] LID - 93 AB - BACKGROUND: This study is aimed to screen out the microRNAs (miRNAs) associated with H(2)O(2) induced oxidative stress in human lens epithelial B3 (HLE-B3) cell lines and investigate their relations with the progression of age-related nuclear cataract. METHODS: H(2)O(2) was used to induce oxidative stress in HLE-B3 cells. A genome-wide expression profiling of miRNAs in HLE-B3 cells was performed to select the differentially expressed miRNAs before and after H(2)O(2) treatment. The selected miRNAs were validated by RT-PCR and fluorescence in situ hybridization (FISH). Clinical specimens were divided into three groups according to the Lens Opacities Classification System III (LOCSIII) and the expression levels of the selected miRNAs were tested by RT-PCR in the three groups. Bioinformatics analyses were applied to predict the target genes of the miRNA hits and construct the miRNA regulatory network. The expression level of MAPK14 was analyzed by Western blot. RESULTS: The H(2)O(2) induced oxidative stress model of HLE-B3 cells was established. Nineteen upregulated and 30 downregulated miRNAs were identified as differentially expressed miRNAs. Seven of the total 49 were validated in the cell model. RT-PCR of the clinical samples showed that the expression levels of miR-34a-5p, miR-630 and miR-335-3p were closely related with the severity of nuclear opacity. The images taken from FISH confirmed the results of RT-PCR. There were 172 target genes of the three miRNAs clustered in the category of response to stress. The regulatory network demonstrated that 23 target genes were co-regulated by multiple miRNAs. MAPK14 was the target gene of three miRNAs and the result were verified by Western blot. CONCLUSION: Up-regulation of miR-34a-5p and miR-630 and down-regulation of miR-335-3p are related with the progression of age-related nuclear cataract and the underlying mechanism awaits further functional research to reveal. FAU - Wang, Song AU - Wang S AD - Department of Ophthalmology, Tangdu Hospital, Fourth Military Medical University, 1 Xinsi Road, Xi'an, Shaanxi, 710038, People's Republic of China. FAU - Guo, Chenjun AU - Guo C AD - Department of Ophthalmology, Tangdu Hospital, Fourth Military Medical University, 1 Xinsi Road, Xi'an, Shaanxi, 710038, People's Republic of China. FAU - Yu, Mengsi AU - Yu M AD - Department of Dermatology, Xijing Hospital, Fourth Military Medical University, 169 West Changle Road, Xi'an, Shaanxi, 710032, People's Republic of China. FAU - Ning, Xiaona AU - Ning X AD - Department of Ophthalmology, Tangdu Hospital, Fourth Military Medical University, 1 Xinsi Road, Xi'an, Shaanxi, 710038, People's Republic of China. FAU - Yan, Bo AU - Yan B AD - The State Key Laboratory of Cancer Biology, Department of Biochemistry and Molecular Biology, Fourth Military Medical University, 169 West Changle Road, Xi'an, Shaanxi, 710032, People's Republic of China. FAU - Zhao, Jing AU - Zhao J AD - The State Key Laboratory of Cancer Biology, Department of Biochemistry and Molecular Biology, Fourth Military Medical University, 169 West Changle Road, Xi'an, Shaanxi, 710032, People's Republic of China. FAU - Yang, Angang AU - Yang A AD - The State Key Laboratory of Cancer Biology, Department of Biochemistry and Molecular Biology, Fourth Military Medical University, 169 West Changle Road, Xi'an, Shaanxi, 710032, People's Republic of China. FAU - Yan, Hong AU - Yan H AD - Department of Ophthalmology, Tangdu Hospital, Fourth Military Medical University, 1 Xinsi Road, Xi'an, Shaanxi, 710038, People's Republic of China. zeratulws@126.com. AD - Chongqing Key Laboratory of Ophthalmology and Chongqing Eye Institute, The First Affiliated Hospital of Chongqing Medical University, 1 Youyi Road, Chongqing, 400016, People's Republic of China. zeratulws@126.com. LA - eng GR - 81370997/the National Natural Science Foundation of China/ PT - Journal Article DEP - 20180413 PL - England TA - BMC Ophthalmol JT - BMC ophthalmology JID - 100967802 RN - 0 (MicroRNAs) RN - BBX060AN9V (Hydrogen Peroxide) RN - EC 2.7.11.24 (Mitogen-Activated Protein Kinase 14) RN - Cataract, Age-Related Nuclear SB - IM MH - Blotting, Western MH - Cataract/genetics/*metabolism MH - Cell Line MH - Epithelial Cells/metabolism MH - Gene Expression Profiling MH - Humans MH - Hydrogen Peroxide/pharmacology MH - Lens, Crystalline/*metabolism MH - MicroRNAs MH - Mitogen-Activated Protein Kinase 14/metabolism MH - Oxidative Stress/*physiology MH - Real-Time Polymerase Chain Reaction PMC - PMC5899325 OTO - NOTNLM OT - Age-related nuclear cataract OT - Bioinformatics analysis OT - Oxidative stress OT - microRNA COIS- ETHICS APPROVAL AND CONSENT TO PARTICIPATE: Verbal consent was obtained from each patient following an explanation of the surgery procedure and the purpose of this research. The Moral and Ethical Committee of the Fourth Military Medical University approved the verbal consent and the research program. The research was performed in accordance with the tenets of the Declaration of Helsinki. COMPETING INTERESTS: The authors declare that they have no competing interests. PUBLISHER'S NOTE: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. EDAT- 2018/04/15 06:00 MHDA- 2018/05/01 06:00 PMCR- 2018/04/13 CRDT- 2018/04/15 06:00 PHST- 2017/10/20 00:00 [received] PHST- 2018/04/03 00:00 [accepted] PHST- 2018/04/15 06:00 [entrez] PHST- 2018/04/15 06:00 [pubmed] PHST- 2018/05/01 06:00 [medline] PHST- 2018/04/13 00:00 [pmc-release] AID - 10.1186/s12886-018-0766-6 [pii] AID - 766 [pii] AID - 10.1186/s12886-018-0766-6 [doi] PST - epublish SO - BMC Ophthalmol. 2018 Apr 13;18(1):93. doi: 10.1186/s12886-018-0766-6.