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长链非编码 RNA XIST 通过上调 ETS1 抑制 miR-377-3p 来阻碍 Th17 细胞分化。

LNCRNA XIST Inhibits miR-377-3p to Hinder Th17 Cell Differentiation through Upregulating ETS1.

机构信息

Beijing IROT Key Laboratory, The 8th Medical Center of Chinese PLA General Hospital, 100091 Beijing City, China.

Department of Urology, The 8th Medical Center of Chinese PLA General Hospital, 100091 Beijing City, China.

出版信息

Comput Intell Neurosci. 2022 Jun 14;2022:6545834. doi: 10.1155/2022/6545834. eCollection 2022.

DOI:10.1155/2022/6545834
PMID:35747716
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9213139/
Abstract

BACKGROUND

Th17 cell differentiation is involved in the development and progression of many diseases, such as rheumatoid arthritis and systemic lupus erythematosus. Present study mainly focused on the role of LINC-XIST in Th17 cell differentiation.

METHODS

The naïve CD4+ T cells were isolated from human whole blood. Cells were cultured under Th17 cell-polarizing condition for 6 days. The expression of LINC-XIST and miR-153-3p was measured by qPCR. The relationship between LINC-XIST, miR-153-3p, and ETS1 was predicted by TargetScan website and authenticated by luciferase reporter assay. ELISA assays were conducted to evaluate the IL-17 concentration. Western blot was utilized to measure the protein expression of ETS1. Th17 cell frequency was examined by flow cytometry.

RESULTS

The expression of XIST markedly decreased and miR-153-3p expression markedly increased with Th17 cell differentiation. The mRNA expression of IL-17, IL-17 concentration, and Th17 cell frequency were observably decreased in overexpressed LINC-XIST group. Luciferase reporter assay authenticated that miR-153-5p was directly regulated by LINC-XIST. miR-153-3p inhibitor observably decreased IL-17 concentration, mRNA expression of IL-17, and Th17 cell frequency while si-XIST reversed this impact. ETS1 was confirmed to be regulated by miR-153-5p via luciferase reporter assay. In addition, ETS1 markedly decreased IL-17 mRNA expression, IL-17 concentration, and Th17 cell frequency while miR-153-5p mimic reversed this impact.

CONCLUSION

LNCRNA XIST inhibited miR-377-3p to hinder Th17 cell differentiation through upregulating ETS1.

摘要

背景

Th17 细胞分化参与许多疾病的发展和进展，如类风湿关节炎和系统性红斑狼疮。本研究主要关注 LINC-XIST 在 Th17 细胞分化中的作用。

方法

从人全血中分离出幼稚 CD4+T 细胞。细胞在 Th17 细胞极化条件下培养 6 天。通过 qPCR 测量 LINC-XIST 和 miR-153-3p 的表达。TargetScan 网站预测 LINC-XIST、miR-153-3p 和 ETS1 之间的关系，并通过荧光素酶报告基因检测进行验证。ELISA 检测试剂盒用于评估 IL-17 浓度。Western blot 用于测量 ETS1 的蛋白表达。流式细胞术用于检测 Th17 细胞频率。

结果

随着 Th17 细胞分化，XIST 的表达显著降低，miR-153-3p 的表达显著增加。过表达 LINC-XIST 组的 IL-17mRNA 表达、IL-17 浓度和 Th17 细胞频率明显降低。荧光素酶报告基因检测证实 miR-153-5p 可直接受 LINC-XIST 调控。miR-153-3p 抑制剂显著降低了 IL-17 浓度、IL-17mRNA 表达和 Th17 细胞频率，而 si-XIST 逆转了这一影响。荧光素酶报告基因检测证实 ETS1 受 miR-153-5p 通过调控。此外，ETS1 显著降低了 IL-17mRNA 表达、IL-17 浓度和 Th17 细胞频率，而 miR-153-5p 模拟物逆转了这一影响。

结论

LncRNA XIST 通过上调 ETS1 抑制 miR-377-3p 抑制 Th17 细胞分化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928b/9213139/b93948ef8b67/CIN2022-6545834.006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928b/9213139/b6b4b8957639/CIN2022-6545834.001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928b/9213139/ad36403b4c0f/CIN2022-6545834.002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928b/9213139/af57d7ebe111/CIN2022-6545834.003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928b/9213139/998a9143e709/CIN2022-6545834.004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928b/9213139/3f7f52386836/CIN2022-6545834.005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928b/9213139/b93948ef8b67/CIN2022-6545834.006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928b/9213139/b6b4b8957639/CIN2022-6545834.001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928b/9213139/ad36403b4c0f/CIN2022-6545834.002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928b/9213139/af57d7ebe111/CIN2022-6545834.003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928b/9213139/998a9143e709/CIN2022-6545834.004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928b/9213139/3f7f52386836/CIN2022-6545834.005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/928b/9213139/b93948ef8b67/CIN2022-6545834.006.jpg

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