Purpose
Graves' ophthalmopathy (GO) is a visually debilitating and cosmetically disfiguring orbital disorder, characterized by the remodeling of extraocular muscles (EOMs). This study aimed to investigate the role of long non-coding RNAs (lncRNAs) in the EOMs of GO.
Methods
LncRNA microarray analysis was performed on EOM tissues sampled from GO and concomitant esotropia patients. LncRNA ENST00000581911 was identified and subjected to bioinformatics analysis. High-throughput RNA sequencing, CCK-8, and ELISA were used to investigate the regulatory function of ENST00000581911 in vitro. Furthermore, RNA pull-down, LC-MS/MS, and WB were applied to identify the RNA-binding protein (RBP) interacting with ENST00000581911.
Results
A total of 1,261 lncRNAs were identified as differentially expressed in the EOMs of GO, with 648 up-regulated and 613 down-regulated. Among these lncRNAs, the up-regulated lncRNA ENST00000581911 exhibited the highest expression level, as validated by qRT-PCR. Functional analysis demonstrated that ENST00000581911 might be involved in inflammatory response, regulation of muscle contraction, and amino sugar and nucleotide sugar metabolism. RNA sequencing showed that ENST00000581911 might play a regulatory role in DNA replication, extracellular matrix and cell cycle of orbital fibroblasts (OFs). CCK-8 and ELISA assays showed that ENST00000581911 enhanced the cell viability and promoted hyaluronic acid secretion of OFs. Additionally, KHSRP was identified as the RBP of ENST00000581911, and they interacted with each other to regulate OF function.
Conclusions
This study provides novel insights into the role of lncRNAs, particularly ENST00000581911, in the pathogenesis of EOM remodeling in GO. ENST00000581911 may serve as a potential therapeutic target of GO. Less...