胃肠间质瘤中差异表达长链非编码RNA和mRNA的筛选鉴定及功能分析

Screening,Identification and Functional Analysis of Differentially Expressed Long Non-coding RNAs and mRNAs in Gastrointestinal Stromal Tumor

[目的]应用生物信息学技术鉴定胃肠间质瘤(GISTs)中差异表达的长链非编码RNA(lncRNA)和m RNA,并对其功能进行初步分析。[方法]在GEO数据库检索GISTs表达情况,并进行主成分分析、靶分子鉴定、GO和KEGG富集分析等;qPCR法检测10例临床GISTs组织及正常黏膜组织中部分差异分子的表达情况,对数据库结果进行验证。[结果]热图分析显示,差异表达在GISTs组织及正常黏膜分析识别出293个差异表达的lncRNA和452个差异表达的mRNA。应用皮尔森相关分析进行trans调控筛选,分析相关性水平,发现前30%的lncRNA和mRNA存在互作对,这些差异表达的lncRNA和mRNA通过靶分子作用参与了GISTs的细胞增殖及细胞迁移等生物学功能,其中lncRNA VCAN-AS1和AMPD3 mRNA在GISTs中表达高于正常黏膜,且可能存在相互作用。对lncRNA VCAN-AS1和AMPD3 mRNA在GISTs中表达进行验证显示,lncRNA VCAN-AS1和AMPD3 mRNA在GISTs中表达(2.762±1.593,1.545±1.162)高于正常黏膜(0.506±0.183,0.702±0.155)(t=4.449,P <0.001;t=2.274,P=0.035),GISTs组织中lncRNA VCAN-AS1与AMPD3 mRNA表达呈正相关(r=0.9273,P<0.001),与生物信息学分析结果一致。[结论] GISTs中存在lncRNA和mRNA的差异表达,lncRNA VCAN-AS1和AMPD3可能通过lncRNA-mRNA调控机制参与了GISTs的发生和进展。

[Objective] To screen,identify and analyze the differentially expressed long non-coding RNA(lnc RNA) and m RNA in gastrointestinal stromal tumors(GISTs) by applying bioinformatics. [Methods]GEO database was used for investigating GISTs expression;and the principal component analysis,target molecular identification,GO and KEGG enrichment analysis were performed. The q PCR method was used to detect the expression of differentially expressed molecules in 10 cases of clinical GISTs tissues and normal mucosa tissues adjacent to cancer to verify the database results. [Results] Heat map analysis showed that 293 differentially expressed lnc RNAs and 452 differentially expressed m RNAs were identified. Pearson correlation analysis was used for trans regulation screening to analyze the interaction between lnc RNA and m RNA with the correlation level of top 30%. These differentially expressed lnc RNA and m RNA were involved in the biological functions of GISTs such as cell proliferation and cell migration through target molecules. Lnc RNA VCAN-AS1 and AMPD3 m RNA expression increased in GISTs and there might be an interaction between them. The expression of lnc RNA VCAN-AS1 and AMPD3 m RNA in GISTs was verified in GISTs tissue. The expression of lnc RNA VCAN-AS1 and AMPD3 m RNA in GISTs(2.762±1.593,1.545±1.162) was higher than that in normal mucosa(0.506±0.183,0.702±0.155)(t=4.449,P <0.001;t =2.274,P =0.035),and lnc RNA VCAN-AS1 was positively correlated with AMPD3 m RNA expression in GISTs tissues(r=0.9273,P<0.001),which were consistent with the results of bioinformatics analysis. [Conclusion] There are differential expressions of lnc RNA and m RNA in GISTs,lnc RNA VCAN-AS1 and AMPD3 m RNA may participate in the occurrence and progress of GISTs through lncRNA-mRNA regulation mechanism.