冠状动脉支架内再狭窄相关miRNA的表达谱分析和意义

MicroRNA profiling and significance in coronary in-stent restenosis

目的筛选出支架内再狭窄(ISR)患者中特异性表达的miRNA,并用生物信息学方法分析其意义。方法通过miRNA基因芯片筛选ISR患者血浆miRNA的表达谱差异,以筛选出与ISR相关的miRNA;通过实时荧光定量聚合酶链反应(Real-time PCR)对miRNA的相对表达量进行验证;通过生物信息学技术对miRNA靶基因进行预测,并对miRNA的下游靶基因网络进行数据挖掘,构建ISR相关miRNA-Gene-Network网络,探讨其在ISR中可能的作用机制。结果经过高通量miRNA芯片检测,筛选获得了一批在ISR患者中差异表达的miRNA,其中43个基因表达下调,10个基因表达上调。通过生物信息学方法及Real-time PCR对基因芯片结果进行了验证,获得了一批与ISR相关的miRNA及靶基因,并构建了miRNA与靶基因的调控网络miRNA-Gene-Network。Real-time PCR定量检测发现,与non-ISR组比较,ISR组miR-126明显下降(0.507±0.131比1.427±0.337,P<0.05)。结论 miRNA在ISR患者中存在差异性表达,这些miRNA及靶基因可能与ISR的发生有关,miR-126水平下降倾向于发生ISR。

Aim To screen out the specific expression of miRNA in ISR,and its significance was analyzed by bioinformatics method. Methods The expression profiles of miRNA in ISR patients were screened by miRNA gene chip to screen out ISR related miRNA. The relative expression of miRNA and ISR was verfied by Real-time PCR. miRNA target genes were predicted by bioinformatics,and data mining of the downstream target genes of miRNA was carried out,and the ISR related miRNA-Gene-Network was constructed,the possible mechanism of ISR was discussed. Results The difference expression of miRNA in ISR patients was detected by high throughput miRNA microarray,43 genes were down regulated and 10 genes were up-regulated. Bioinformatics and Real-time PCR were used to verfy the result of microarray. A number of miRNA and target genes related to ISR were obtained,and the regulatory networks of miRNA and target genes were constructed miRNA-Gene-Network. Quantitative detection of Real-time PCR quantitative detection showed that miR-126 in group ISR was significantly lower than that in group non-ISR（ 0.507±0.131 vs. 1.427±0.337,P〈0. 05）. Conclusion The expression of miRNA in ISR patients is different from no-ISR,these miRNA and target genes may be related to the occurrence of ISR,the decrease of miR-126 level tended to occur in ISR.