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通过微阵列鉴定的克罗恩病患者血浆长链非编码RNA表达谱

Plasma long noncoding RNA expression profile identified by microarray in patients with Crohn's disease.

作者信息

Chen Dong, Liu Jiang, Zhao Hui-Ying, Chen Yi-Peng, Xiang Zun, Jin Xi

机构信息

Dong Chen, Hui-Ying Zhao, Department of Colorectal Surgery, the First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, Zhejiang Province, China.

出版信息

World J Gastroenterol. 2016 May 21;22(19):4716-31. doi: 10.3748/wjg.v22.i19.4716.

Abstract

AIM

To investigate the expression pattern of plasma long noncoding RNAs (lncRNAs) in Chrohn's disease (CD) patients.

METHODS

Microarray screening and qRT-PCR verification of lncRNAs and mRNAs were performed in CD and control subjects, followed by hierarchy clustering, GO and KEGG pathway analyses. Significantly dysregulated lncRNAs were categorized into subgroups of antisense lncRNAs, enhancer lncRNAs and lincRNAs. To predict the regulatory effect of lncRNAs on mRNAs, a CNC network analysis was performed and cross linked with significantly changed lncRNAs. The overlapping lncRNAs were randomly selected and verified by qRT-PCR in a larger cohort.

RESULTS

Initially, there were 1211 up-regulated and 777 down-regulated lncRNAs as well as 1020 up-regulated and 953 down-regulated mRNAs after microarray analysis; a heat map based on these results showed good categorization into the CD and control groups. GUSBP2 and AF113016 had the highest fold change of the up- and down-regulated lncRNAs, whereas TBC1D17 and CCL3L3 had the highest fold change of the up- and down-regulated mRNAs. Six (SNX1, CYFIP2, CD6, CMTM8, STAT4 and IGFBP7) of 10 mRNAs and 8 (NR_033913, NR_038218, NR_036512, NR_049759, NR_033951, NR_045408, NR_038377 and NR_039976) of 14 lncRNAs showed the same change trends on the microarray and qRT-PCR results with statistical significance. Based on the qRT-PCR verified mRNAs, 1358 potential lncRNAs with 2697 positive correlations and 2287 negative correlations were predicted by the CNC network.

CONCLUSION

The plasma lncRNAs profiles provide preliminary data for the non-invasive diagnosis of CD and a resource for further specific lncRNA-mRNA pathway exploration.

摘要

目的

研究克罗恩病(CD)患者血浆长链非编码RNA(lncRNA)的表达模式。

方法

对CD患者和对照者进行lncRNA和mRNA的微阵列筛选及qRT-PCR验证,随后进行层次聚类、GO和KEGG通路分析。将显著失调的lncRNA分为反义lncRNA、增强子lncRNA和长链基因间非编码RNA(lincRNA)亚组。为预测lncRNA对mRNA的调控作用,进行了CNC网络分析,并与显著变化的lncRNA进行交联。随机选择重叠的lncRNA,在更大的队列中通过qRT-PCR进行验证。

结果

最初,微阵列分析后有1211个lncRNA上调、777个lncRNA下调,以及1020个mRNA上调、953个mRNA下调;基于这些结果的热图显示CD组和对照组分类良好。GUSBP2和AF113016在lncRNA上调和下调中倍数变化最高,而TBC1D17和CCL3L3在mRNA上调和下调中倍数变化最高。10个mRNA中的6个(SNX1、CYFIP2、CD6、CMTM8、STAT4和IGFBP7)和14个lncRNA中的8个(NR_033913、NR_038218、NR_036512、NR_049759、NR_033951、NR_045408、NR_038377和NR_039976)在微阵列和qRT-PCR结果上显示相同变化趋势且具有统计学意义。基于qRT-PCR验证的mRNA,通过CNC网络预测了1358个潜在lncRNA,其中2697个具有正相关,2287个具有负相关。

结论

血浆lncRNA谱为CD的非侵入性诊断提供了初步数据,并为进一步探索特定的lncRNA-mRNA通路提供了资源。

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