Deduction of Novel Genes Potentially Involved in the Effects of Very Low Dose Atropine (0.003%) Treatment on Corneal Epithelial Cells Using Next-Generation Sequencing and Bioinformatics Approaches
Deduction of Novel Genes Potentially Involved in the Effects of Very Low Dose Atropine (0.003%) Treatment on Corneal Epithelial Cells Using Next-Generation Sequencing and Bioinformatics Approaches
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DOI：
10.3390/medicina55090589
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发表时间：
2019-09-01
影响因子：
2.6
通讯作者：
Kuo, Po-Lin
中科院分区：
文献类型：
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作者：
Chang, Wei-An;Hsiao, Yu-Ting;Kuo, Po-Lin
背景和目的：阿托品是一种非选择性的毒蕈碱拮抗剂，已被用于预防儿童近视的恶化。不同浓度的阿托品用于近视，范围为0.01%至1.0%。但不同剂量的阿托品对角膜仍有潜在的毒性作用。在这里，我们提出了一项研究，调查新的基因可能涉及的影响非常低剂量阿托品治疗（0.003%）角膜上皮细胞使用下一代测序（NGS）和生物信息学方法。材料与方法：用0.003%阿托品处理人角膜上皮细胞，培养至汇合，并提取RNA用于对照和阿托品处理的角膜上皮细胞之间的mRNA和microRNA（miRNA）的差异表达谱。差异表达基因的功能富集分析使用两个生物信息学数据库进行，包括数据库注释，可视化和集成发现（大卫）和Incubity（R）途径分析（IPA）。此外，潜在的miRNA-mRNA相互作用参与阿托品处理的角膜上皮细胞进行了预测和验证，使用不同的miRNA目标预测数据库。结果如下：结果表明，0.003%阿托品可能通过Ras和蛋白激酶A信号通路抑制角膜上皮细胞凋亡。我们还通过TargetScan和miRDB数据库验证了可能的miRNA调控。经验证，hsa-miR-651- 3 p-EPHA 7、hsa-miR-3148-TMEM 108和hsa-miR-874- 5 p-TBX 6可能是参与用0.003%阿托品处理的角膜上皮细胞的miRNA调控。结论：这些发现可能有助于新的见解，治疗策略的角膜与0.003%阿托品。
Background and Objectives: Atropine is a nonselective muscarinic antagonist which has been used to prevent worsening of myopia in children. Different concentrations of atropine were used for myopia, ranging from 0.01% to 1.0%. However, there are still potential toxicity of different doses of atropine to the cornea. Here, we present a study of investigating novel genes potentially involved in the effects of very low dose atropine treatment (0.003%) on corneal epithelial cells using next-generation sequencing (NGS) and bioinformatics approaches. Materials and Methods: Human corneal epithelial cells were treated with 0.003% atropine, cultured until confluence, and RNA extracted for differential expression profiling of mRNA and microRNA (miRNA) between control and atropine-treated corneal epithelial cells. The functional enrichment analysis for differentially expressed genes was performed using two bioinformatics databases, including Database for Annotation, Visualization and Integrated Discovery (DAVID) and Ingenuity (R) Pathway Analysis (IPA). In addition, potential miRNA-mRNA interactions involved in atropine-treated corneal epithelial cells were predicted and validated using different miRNA target prediction databases. Results: Our results showed 0.003% atropine might suppress the apoptosis of corneal epithelial cells, potentially through Ras and protein kinase A signaling pathways. We also validated the possible miRNA regulations by using TargetScan and miRDB databases. Hsa-miR-651-3p-EPHA7, hsa-miR-3148-TMEM108 and hsa-miR-874-5p-TBX6 were validated as possible miRNA regulations involved in corneal epithelial cells treated with 0.003% atropine. Conclusions: These findings may contribute novel insights into therapeutic strategies for treating cornea with 0.003% atropine.