Integrated Analysis of Long Non-Coding RNA -mRNA Profile and Validation in Diabetic Cataract

To illustrate the expression profile of lncRNAs and mRNAs in diabetic cataract (DC) and explore their potential role in the pathogenesis of DC. LncRNA and mRNA microarray were conducted using the lens epithelium of DC patients (n = 3) and controls from eye bank (n = 3). Further, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were employed to unveil their underlying biological function and signaling pathway. The co-expression network was constructed to explore the relationship between lncRNAs and mRNAs. Then, lens epithelial cells (LECs) were cultured in vitro under high glucose (HG) and normal glucose (NG) groups to further verified the expression profiles of mRNAs. At last, quantitative RT-PCR (qRT-PCR) was performed to confirm the microarray results for the aberrantly expressed lncRNAs, and to unveil the expression prolife of mRNAs in LECs. Eight thousand three hundred and twenty-six lncRNAs and 3303 mRNAs were dysregulated in DC. GO analysis unveiled that the upregulated mRNAs, highest enrichment score of the GO term belonged to oxidation-reduction process, while the highest for downregulated mRNAs went to protein lipidation. In the CC category, the most significant terms for upregulated and downregulated mRNAs appeared in intracellular part and intrinsic component of membrane, respectively. As to MF category, the most significant term for upregulated mRNAs was protein binding, and for downregulated mRNAs was catecholamine binding. KEGG pathways analysis demonstrated that the major enrichment score of pathways in upregulated mRNAs included Glycolysis/Gluconeogenesis, lysosome, carbon metabolism and AGE-RAGE signaling pathway. For the downregulated mRNAs, the glycosphingolipid biosynthesis-lacto and neolacto series pathway was included. Three lncRNAs (LINC01508, MAFA-AS1, MIAT) and the <i>Sirtuin 2</i> are positively related in the co-expression network. This study provided an overview of the lncRNAs and mRNAs expression, and suggesting that several aberrantly expressed lncRNAs might participate in the pathogenesis of DC.

为阐明糖尿病性白内障（diabetic cataract, DC）患者体内长链非编码RNA（long non-coding RNA, lncRNAs）与信使RNA（messenger RNA, mRNAs）的表达谱，并探讨二者在DC发病机制中的潜在作用。本研究采集3例DC患者与3例眼库来源对照者的晶状体上皮组织，开展了lncRNA与mRNA芯片检测。进一步通过基因本体论（Gene Ontology, GO）与京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes, KEGG）通路分析，揭示二者潜在的生物学功能与信号通路。构建共表达网络，以探究lncRNAs与mRNAs之间的相互关系。随后，体外培养高糖（high glucose, HG）与正常糖（normal glucose, NG）环境下的晶状体上皮细胞（lens epithelial cells, LECs），以进一步验证mRNAs的表达谱。最后，通过实时定量逆转录聚合酶链反应（quantitative reverse transcription polymerase chain reaction, qRT-PCR），验证差异表达lncRNAs的芯片检测结果，并明确LECs中mRNAs的表达谱。DC患者体内共存在8326个差异表达lncRNAs与3303个差异表达mRNAs。GO分析显示，上调mRNAs富集得分最高的GO条目为氧化还原过程，而下调mRNAs富集得分最高的GO条目为蛋白质脂化修饰。在细胞组分（Cellular Component, CC）大类中，上调与下调mRNAs最显著的富集条目分别定位于细胞内区域与膜固有成分。在分子功能（Molecular Function, MF）大类中，上调mRNAs最显著的富集条目为蛋白质结合，而下调mRNAs则为儿茶酚胺结合。KEGG通路分析显示，上调mRNAs主要富集的通路包括糖酵解/糖异生、溶酶体、碳代谢以及晚期糖基化终末产物-晚期糖基化终末产物受体（AGE-RAGE）信号通路。下调mRNAs富集于糖鞘脂生物合成-乳糖系列与新乳糖系列通路。共表达网络中，LINC01508、MAFA-AS1、MIAT这3个lncRNAs与沉默信息调节因子2（Sirtuin 2）呈正相关关系。本研究全面梳理了DC患者体内lncRNAs与mRNAs的表达特征，提示若干差异表达lncRNAs可能参与DC的发病过程。