关于精神分裂症相关潜在 miRNA 在 COMT 基因调控中的作用的体外研究

IF 5.5 2区 医学 Q1 NEUROSCIENCES Molecular Neurobiology Pub Date : 2024-10-01 Epub Date: 2024-03-01 DOI:10.1007/s12035-024-04070-2
Onur Tonk, Pervin Elvan Tokgun, Özge Sarıca Yılmaz, Onur Tokgun, Kubilay Inci, Büşra Çelikkaya, Nuray Altintas
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引用次数: 0

摘要

本研究旨在分析作为精神分裂症潜在候选miRNA的miR-30a-5p、miR-30e-5p和miR-34a-5p与COMT基因可能存在的关联。候选 miRNA 来自 TargetScan 数据库。将 miR-30a-5p、miR-30e-5p 和 miR-34a-5p 模拟物转染到 SH-SY5Y 细胞系中。从转染细胞和 RNA-IP 样品中分离出总 RNA,并进行反转录,以分析 miRNA 和 mRNA。进行RT-qPCR和Western印迹以观察COMT表达水平的变化。进行 RNA-ımmunoprecipitation 以确定模拟转染后的 RNA 蛋白相互作用。研究发现,miR-30a-5p 和 miR-34a-5p 表达后,COMT 基因表达水平显著下降,而 miR-30e-5p 转染后,COMT 基因表达水平显著上升。RNA-IP 数据显示,转染 miR-30a-5p 和 miR-34a-5p 后,Ago2 拉下的 COMT 数量增加。RNA-IP 结果显示,miR-30a-5p 和 miR-34a-5p 是 COMT 基因的直接靶标。
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An In Vitro Study for the Role of Schizophrenia-Related Potential miRNAs in the Regulation of COMT Gene.

This study aimed to analyze the possible association of miR-30a-5p, miR-30e-5p, and miR-34a-5p identified as potential candidate miRNAs in schizophrenia, with the COMT gene. Candidate miRNAs were obtained from the TargetScan database. The SH-SY5Y human neuroblastoma cell line was used as a cellular model for schizophrenia. miR-30a-5p, miR-30e-5p, and miR-34a-5p mimics were transfected into the SH-SY5Y cell line. Total RNA was isolated from transfected cells and RNA-IP samples and reverse transcripted for miRNA and mRNA analysis. RT-qPCR and western blot were performed to observe changes in expression levels of COMT. RNA-ımmunoprecipitation was performed to determine RNA-protein interactions after mimic transfection. In the study, it was observed that COMT gene expression levels decreased significantly after miR-30a-5p and miR-34a-5p expressions, whereas increased significantly as a result of miR-30e-5p transfection. RNA-IP data have shown that the amount of COMT pulled down by Ago2 was increased after miR-30a-5p and miR-34a-5p transfections. RNA-IP results revealed that miR-30a-5p and miR-34a-5p are direct targets for the COMT gene.

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来源期刊
Molecular Neurobiology
Molecular Neurobiology 医学-神经科学
CiteScore
9.00
自引率
2.00%
发文量
480
审稿时长
1 months
期刊介绍: Molecular Neurobiology is an exciting journal for neuroscientists needing to stay in close touch with progress at the forefront of molecular brain research today. It is an especially important periodical for graduate students and "postdocs," specifically designed to synthesize and critically assess research trends for all neuroscientists hoping to stay active at the cutting edge of this dramatically developing area. This journal has proven to be crucial in departmental libraries, serving as essential reading for every committed neuroscientist who is striving to keep abreast of all rapid developments in a forefront field. Most recent significant advances in experimental and clinical neuroscience have been occurring at the molecular level. Until now, there has been no journal devoted to looking closely at this fragmented literature in a critical, coherent fashion. Each submission is thoroughly analyzed by scientists and clinicians internationally renowned for their special competence in the areas treated.
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