芯片技术研究1ncRNA在非IgA系膜增生性肾小球肾炎中的表达  

Expression of long non-coding RNA in patients with non-IgA mesangial proliferative glomerulonephritis

作　　者：丛珊[1] 眭维国[1] 邹贵勉[1] 薛雯[1] 李欢[1] 晏强[1] 陈洁晶[1] 罗雅丹[1] 陈怀周[1] 

机构地区：[1]解放军181医院肾脏科广西代谢性疾病重点实验室

出　　处：《临床肾脏病杂志》2015年第4期234-238,共5页Journal Of Clinical Nephrology

基　　金：广西自然科学基金(NO.2011GXNSFB018105)

摘　　要：目的通过微阵列芯片技术研究非IgA系膜增生肾小球肾炎（non-IgA mesangial proliferative glomerulonephritis，non-IgA MsPGN）组（观察组）及对照组中信使RNA（Message RNA，mRNA）和长链非编码RNA（long non-coding RNA，lncRNA）的差异性表达，从而探索lncRNA在non-IgA MsPGN发病机制中潜在的作用。方法通过单纯随机抽样分别选取4例non-IgAMsPGN患者和2例未患肾病者作为观察组和对照组，分别收集2组的肾皮质组织，提取并鉴定总RNA，然后制备双链cDNA，单色荧光标记后用于芯片杂交。通过Gene Ontology，Pathway分析及mRNA与ln-cRNA基因座位置关联分析，找出与non-IgAMsPGN密切相关的lncRNA。最后，采用半定量逆转录聚合酶链反应（reverse transcription-polymerase chain reaction，RT-PCR）对部分基因的芯片检测结果进行检测，验证基因芯片结果的可靠性。结果经过折叠倍率过滤（fold-change filtering），筛选出有统计学差异（P〈O．05）表达的mRNA转录本4317个与lncRNA转录本3502个。经分析，发现了5个在non-IgAMsPGN发病机制中有着重要潜在作用的lncRNA：AF1180924（相邻编码基因FGG），AK092233（相邻编码基因COL18A1），AKl30579（相邻编码基因CREBBP），AK023598（相邻编码基因LEPR），AK055915（相邻编码基因CDC42EP3），为全面揭示nonqgAMsPGN发病机制提供了重要依据。结论某些lncRNA可潜在调控相关基因，在non-IgAMsPGN的发病机制及发展过程中起重要作用。Objective To study differential expression profile of mRNA and long non-coding RNA （lncRNA） through microarray analysis between non-IgA mesangial proliferative glomerulone- phritis （MsPGN） patients and the controls, and then explore the potential role of lncRNA in the pathogenesis of non-IgA MsPGN. Methods Through simple random sampling, 4 patients with nowlgA MsPGN and 2 controls were selected as disease group and control group, respeetively. Renal cortical tissues from two groups were collected. Total RNA was extracted, quantified and prepared to ds-cD- NA through reverse transcription, ds-cDNA was labeled with NimbleGen one-color DNA labeling kit and used for array hybridization. All experimental data were processed through GO analysis, Pathway analysis and the gene loci correlation analysis of mRNA and lncRNA. Some lncRNAs that were closely related to non-IgA MsPGN were screened out. Finally, part of the array results was detected by PCR to verify the reliability of array test. Results By fold change filtering, 4317 differentially expressed mRNAs and 3502 differentially expressed lncRNAs were screened out. Five lncRNAs were found to play potential roles in the pathogenesis of non-IgA MsPGN： AF1180924 （close to coding gene FGG）,AK092233 （close to coding gene COL18A1 ）, AK130579 （close to coding gene CREBBP）, AK023598 （close to coding gene LEPR）, and AK055915 （close to coding gene CDC42EP3 ）. These results provided an important basis for revealing the patbogenesis of non-IgA MsPGN. Conclusions Some lncRNAs can potentially regulate related genes and plays an important role in the pathogenesis and development of non-IgA MsPGN.

关 键 词：长链非编码RNA 非IgA系膜增生肾小球肾炎 差异性表达 微阵列芯片 

分 类 号：R692.31[医药卫生—泌尿科学]