细颗粒物暴露下人眼角膜上皮细胞中长链非编码 RNA 的表达谱。

Expression profiles of long noncoding RNAs in human corneal epithelial cells exposed to fine particulate matter.

机构信息

Eye Center of the 2nd Affiliated Hospital, School of Medicine, Zhejiang University, Zhejiang Provincial Key Lab of Ophthalmology, Hangzhou, Zhejiang Province, China.

Department of Environmental and Occupational Health, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou, Zhejiang Province, China.

出版信息

Chemosphere. 2022 Jan;287(Pt 1):131955. doi: 10.1016/j.chemosphere.2021.131955. Epub 2021 Aug 21.

DOI:10.1016/j.chemosphere.2021.131955

PMID:34478962

Abstract

PURPOSE

The aim of this study was to investigate the expression profiles of long noncoding RNAs (lncRNAs) in human corneal epithelial cells (HCECs) exposed to fine particulate matter (PM) and to identify potential biological pathways involved in PM-induced toxicity in HCECs.

METHODS

Using RNA sequencing (RNA-seq) and hierarchy clustering analysis, lncRNA expression profiles in PM-treated and untreated HCECs were examined. Gene ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed to predict the role of altered lncRNAs in biological processes and pathways. A quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) assay was conducted to verify the RNA-seq results in HCECs and human corneal epithelial cell sheets.

RESULTS

In total, 65 lncRNAs were altered in the PM-treated HCECs, including 41 upregulated and 24 downregulated lncRNAs. The results of the qRT-PCR assay were consistent with those of the RNA-seq analysis. The expression of two significantly upregulated lncRNAs was confirmed in human corneal epithelial cell sheets. The GO analysis demonstrated that altered lncRNAs in the PM-treated HCECs were significantly enriched in three domains: cellular component, molecular function, and biological process. The KEGG pathway analysis revealed enriched pathways of lncRNA co-expressed mRNAs, including cancer, RNA transport, and Rap1 signaling.

CONCLUSIONS

Our results suggest that lncRNAs are involved in the pathogenesis of PM-induced ocular diseases, exerting their effects through biological processes and pathogenic pathways. Among the altered lncRNAs, RP3-406P24.3 and RP11-285E9.5 may play significant roles in PM-induced ocular surface injury.

摘要

目的

本研究旨在探讨细颗粒物（PM）暴露下人角膜上皮细胞（HCEC）中长链非编码 RNA（lncRNA）的表达谱，并鉴定 PM 诱导的 HCEC 毒性相关的潜在生物学途径。

方法

采用 RNA 测序（RNA-seq）和层次聚类分析，检测 PM 处理和未处理的 HCEC 中的 lncRNA 表达谱。进行基因本体论（GO）富集分析和京都基因与基因组百科全书（KEGG）通路分析，以预测改变的 lncRNA 在生物学过程和途径中的作用。采用实时定量逆转录聚合酶链反应（qRT-PCR）检测验证 HCEC 和人角膜上皮细胞片上的 RNA-seq 结果。

结果

共发现 65 个 lncRNA 在 PM 处理的 HCEC 中发生改变，包括 41 个上调和 24 个下调的 lncRNA。qRT-PCR 检测结果与 RNA-seq 分析结果一致。在人角膜上皮细胞片上确认了两个显著上调的 lncRNA 的表达。GO 分析表明，PM 处理的 HCEC 中改变的 lncRNA 在三个域中显著富集：细胞成分、分子功能和生物学过程。KEGG 通路分析显示 lncRNA 共表达 mRNAs 的富集途径，包括癌症、RNA 转运和 Rap1 信号通路。