血清microRNA表达谱对原发性胆汁性肝硬化的诊断价值

A pilot study of serum microRNA profiles as a novel biomarker for primary biliary cirrhosis

目的:寻找原发性胆汁性肝硬化(primary biliary cirrhosis,PBC)患者血清中差异表达的microRNAs。方法:首先,将PBC组和健康对照组血清标本各3例各自混合,采用Illumina GAⅡx高通量测序技术分析上述2组样本,筛选出PBC患者与健康对照者表达差异有统计学意义的miRNAs;其次,采用实时荧光定量PCR(qRT-PCR)对差异表达的miRNAs在扩大标本中进行验证。最后,预测miRNAs的靶基因并进一步分析其作用。结果:高通量测序技术筛选出143条miRNAs表达差异有统计学意义,其中,22条差异倍数>2倍。经过qRT-PCR验证,miR-122,miR-141,miR-34a在PBC组中的表达水平明显高于对照组,与测序结果表达趋势一致。靶基因预测结果显示,在细胞核和细胞质中,miRNAs主要通过与蛋白质结合发挥信号转导的作用。结论:PBC患者存在差异表达的miRNAs,且其对PBC的诊断有一定的参考价值。

Objective:To screen serum microRNA profiles by high throughput sequencing technology method as diagnostic markers for primary biliary cirrhosis (PBC).Methods:First,we employed Illumina GA Ⅱx deep sequencing for the initial screening to indicate the read numbers of microRNAs expression in serum pool of 3 PBC and 3 healthy controls,respectively.Second,qRT-PCR validation study was conducted in more samples.Finally,computer analysis was conducted to predict target genes and biological functions.Results:Twenty-two miRNAs,whose fold change were greater than 2,were filtered out by high throughput sequencing technology.After qRT-PCR validation,the serum expression levels of miR-122,miR-34a and miR-141 were significantly higher in PBC,consistent with the sequencing results.Target genes of prediction results showed that microRNAs played a role in signal transduction,in the nucleus and cytoplasm,through combined with protein.Conclusion:The distinguishing expressed microRNAs had certain reference value on the diagnosis of PBC.