miR-9 和 miR-193b 在人 Th17 细胞分化中的上调。
Upregulation of miR-9 and miR-193b over human Th17 cell differentiation.
机构信息
Department of Animal Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.
Department of Cell and Molecular Biology and Microbiology, Faculty of Biological Science and Technology, University of Isfahan, Isfahan, Iran.
出版信息
Mol Genet Genomic Med. 2020 Dec;8(12):e1538. doi: 10.1002/mgg3.1538. Epub 2020 Oct 31.
BACKGROUND
Th17 cells are a newly discovered subset of CD4 T cells known as key participants in various immune responses and inflammatory conditions including autoimmune diseases. Mi(cro)RNAs are a family of non-coding RNAs that regulate numerous critical immune functions. Immuno-miRNAs modulate cell biological processes in T cells, such as differentiation and function of Th17 cells. The aim of the present study is to investigate the expression of miR-9-5p, miR-193b-3p, and autoimmunity-related genes during human Th17 cells differentiation.
METHODS
Human naïve CD4 T cells were purified from peripheral blood mononuclear cells (PBMCs) by magnetic cell sorting system (MACS) and their purity was checked by flow-cytometric analysis. Naïve CD4 T cells were cultured under Th17-polarizing condition for 6 days. IL- 17 secretion was determined by means of enzyme-linked immunosorbent assay (ELISA). Next, the expression levels of miRNAs and putative targets genes were assessed by qRT-PCR at different time points of differentiation.
RESULTS
Our result showed dramatic downregulation of TCF7, MAP3K1, ENTPD1, and NT5E genes during human Th17 differentiation. Polarization also had a significant inducible effect on the expression of miR-9 and miR-193b over differentiation of human Th17 cells. According to our results, miR-9-5p and miR-193b-3p may contribute to Th17 differentiation probably by inhibiting the expression of negative regulators of Th17 differentiation.
CONCLUSION
This study confirmed deregulation of TCF7, MAP3K1, ENTPD1, and NT5E genes in Th17 differentiation process and introduced miR-9 and miR-193b as Th17 cell-associated miRNAs, making them good candidates for further investigations.
背景
Th17 细胞是一种新发现的 CD4 T 细胞亚群，被认为是各种免疫反应和炎症状态（包括自身免疫性疾病）的关键参与者。mi(cro)RNAs 是一类非编码 RNA，调节着许多关键的免疫功能。免疫 microRNAs 调节 T 细胞中的细胞生物学过程，如 Th17 细胞的分化和功能。本研究旨在探讨 miR-9-5p、miR-193b-3p 和自身免疫相关基因在人 Th17 细胞分化过程中的表达。
方法
通过磁珠细胞分选系统（MACS）从外周血单核细胞（PBMCs）中纯化人幼稚 CD4 T 细胞，并通过流式细胞术分析检查其纯度。将幼稚 CD4 T 细胞在 Th17 极化条件下培养 6 天。通过酶联免疫吸附试验（ELISA）测定 IL-17 的分泌。然后，在分化的不同时间点通过 qRT-PCR 评估 microRNAs 和假定靶基因的表达水平。
结果
我们的结果显示，在人 Th17 分化过程中，TCF7、MAP3K1、ENTPD1 和 NT5E 基因表达显著下调。极化对 miR-9 和 miR-193b 的表达也有显著的诱导作用，超过了人 Th17 细胞的分化。根据我们的结果，miR-9-5p 和 miR-193b-3p 可能通过抑制 Th17 分化的负调控因子的表达来促进 Th17 分化。
结论
本研究证实了 TCF7、MAP3K1、ENTPD1 和 NT5E 基因在 Th17 分化过程中的失调，并介绍了 miR-9 和 miR-193b 作为与 Th17 细胞相关的 microRNAs，使其成为进一步研究的良好候选者。
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