LncRNA-miRNA-mRNA expression variation profile in the urine of calcium oxalate stone patients
LncRNA-miRNA-mRNA expression variation profile in the urine of calcium oxalate stone patients
复制标题
草酸钙结石患者尿液中LncRNA-miRNA-mRNA表达变化谱
DOI：
10.1186/s12920-019-0502-y
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发表时间：
2019-04-29
影响因子：
2.7
通讯作者：
Wu, Wenqi
中科院分区：
文献类型：
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作者：
Liang, Xiongfa;Lai, Yongchang;Wu, Wenqi
目的探讨草酸钙（CaOx）患者尿液中长链非编码RNA（lncRNA）、微小RNA（miRNA）和信使RNA（mRNA）的表达谱及其生物学功能。从尿液中提取总RNA后，应用miRNA、mRNA和lncRNA芯片检测其表达变化。进行基因本体（GO）富集分析和京都基因和基因组百科全书（KEGG）途径分析以揭示失调的lncRNA相关竞争性内源RNA（ceRNA）网络的基因功能。采用实时荧光定量PCR技术对草酸钠（NaOx）处理的HK-2细胞进行检测，进一步筛选出差异表达的RNA。结果与正常人相比，CaOx患者尿液中差异表达的miRNA共有9个，其中mRNA 883个，lncRNA 1002个。GO分析显示，ceRNA网络中的大部分mRNA在呼吸爆发、线粒体自噬调节和蛋白激酶调节活性方面富集。这些与ceRNA网络相关的基因的KEGG通路分析突出了它们在戊糖磷酸通路、乙醛酸和二羧酸代谢以及Janus激酶/信号转导子和转录激活子（JAK-STAT）信号通路中的关键作用。五种miRNAs（miR-6796-3p、miR-30d-5p、miR-3192-3p、miR-518b和miR-6776-3p），四种mRNA（NT 5E、CDH 4、CLEC 14 A、CCNL 1）和六种lncRNA（lnc-TIGD1L2-3，lnc-KIN-1，lnc-FAM72B-4，lnc-EVI5L-1，lnc-SERPINI1-2，lnc-MB-6）的差异表达与基因芯片结果一致。mRNA和lncRNA可能与信号传导途径的许多变化或代谢和激酶活性的调节有关，为尿石症的早期诊断提供潜在的生物标志物，为进一步研究尿石症的发病机制提供新的依据。
BackgroundTo explore long-non-coding RNA (lncRNA), microRNA (miRNA) and messenger RNA (mRNA) expression profiles and their biological functions in the urine samples in calcium oxalate (CaOx) patients.MethodsFive CaOx kidney stone patients were recruited in CaOx stone group and six healthy people were included as control group, whose midstream morning urine was collected before the patients were given any medicine on admission. After total RNA was extracted from urine, microarray of miRNA, mRNA and lncRNA were applied to explore their expression variation. Gene ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed to reveal the gene functions of the dysregulated lncRNA-associated competing endogenous RNA (ceRNA) network. Quantitative real-time PCR were performed on HK-2 cells treated with sodium oxalate (NaOx) to further screen out the differentially expression profiles of these RNAs.ResultsA total of nine miRNAs, 883 mRNAs and 1002 lncRNAs were differentially expressed in urine of CaOx patients compared with normal population. GO analysis revealed that most of mRNAs from ceRNA network were enriched in terms of respiratory burst, regulation of mitophagy, and protein kinase regulator activity. KEGG pathway analysis of these genes related to ceRNA network highlight their critical role in pentose phosphate pathway, glyoxylate and dicarboxylate metabolism, and Janus kinase/signal transducer and activator of transcription (JAK-STAT) signaling pathway. Five miRNAs (miR-6796-3p, miR-30d-5p, miR-3192–3p, miR-518b and miR-6776-3p), four mRNAs (NT5E, CDH4, CLEC14A, CCNL1) and six lncRNAs (lnc-TIGD1L2–3, lnc-KIN-1, lnc-FAM72B-4, lnc-EVI5L-1, lnc-SERPINI1–2, lnc-MB-6) from the HK-2 cells induced by NaOx were consistent with the expression changes of microarray results.ConclusionThe differential expressed miRNAs, mRNAs and lncRNAs may be associated with numerous variations of the signaling pathways or regulation of metabolism and kinase activity, providing potential biomarkers for early diagnosis of urolithiasis and new basis for further research of urolithiasis mechanism.