Identification of endogenous controls for analyzing serum exosomal miRNA in patients with hepatitis B or hepatocellular carcinoma.

Identification of endogenous controls for analyzing serum exosomal miRNA in patients with hepatitis B or hepatocellular carcinoma.
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鉴定用于分析乙型肝炎或肝细胞癌患者血清外泌体 miRNA 的内源对照

DOI:
10.1155/2015/893594
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发表时间:
2015
期刊:
影响因子:
--
通讯作者:
Pu X
Pu X
中科院分区:
医学4区
文献类型:
--
作者:
Li Y;Zhang L;Liu F;Xiang G;Jiang D;Pu X

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血清外泌体microRNA(miRNAs)作为诊断癌症的潜在生物标志物已受到相当大的关注。用于测量miRNA转录水平的典型技术是逆转录定量聚合酶链反应(RT-qPCR)。验证RT-qPCR数据的一个先决条件是相对于稳定表达的内源性参考基因进行适当的标准化。然而,尚未鉴定出满足外泌体miRNA的对照基因的所有标准的基因。为了找到外泌体miRNAs的控制基因,我们评估了11个已知的参考基因在循环外泌体中的表达稳定性。在这项研究中,我们发现miR-221,miR-191,let-7a,miR-181 a和miR-26 a的组合可以成为使肝脏特异性miRNA表达标准化的最佳基因参考集。这种组合增强了相对定量分析的稳健性。这些发现强调了在定量靶miRNA之前验证参考基因的重要性。此外,我们的发现将改善监测肝炎进展的研究,并有助于发现诊断早期HCC的非侵入性生物标志物。
Serum exosomal microRNAs (miRNAs) have received considerable attention as potential biomarkers for diagnosing cancer. The canonical technique for measuring miRNA transcript levels is reverse transcription quantitative polymerase chain reaction (RT-qPCR). One prerequisite for validating RT-qPCR data is proper normalization with respect to stably expressed endogenous reference genes. However, genes that meet all of the criteria of a control gene for exosomal miRNAs have not yet been identified. To find out the control gene for exosomal miRNAs, we evaluated the expression stability of 11 well-known reference genes in circulating exosomes. In this study, we found that the combination of miR-221, miR-191, let-7a, miR-181a, and miR-26a can be an optimal gene reference set for normalizing the expression of liver-specific miRNAs. This combination enhanced the robustness of the relative quantification analyses. These findings highlight the importance of validating reference genes before quantifying target miRNAs. Furthermore, our findings will improve studies that monitor hepatitis progression and will aid in the discovery of noninvasive biomarkers to diagnose early stage HCC.
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