Identification of reference genes for circulating microRNA analysis in colorectal cancer.
Identification of reference genes for circulating microRNA analysis in colorectal cancer.
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结直肠癌循环 microRNA 分析参考基因的鉴定
DOI：
10.1038/srep35611
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发表时间：
2016-10-19
影响因子：
4.6
通讯作者：
Gou D
中科院分区：
文献类型：
--
作者：
Niu Y;Wu Y;Huang J;Li Q;Kang K;Qu J;Li F;Gou D
定量实时聚合酶链式反应(QPCR)是目前最常用的检测microRNAs(MiRNAs)表达水平的方法，其基础是对内源性参照进行归一化。尽管循环中的miRNAs被认为是潜在的非侵入性疾病生物标志物，但到目前为止还没有关于参照miRNAs用于结直肠癌正常化的研究。在这项研究中，我们试图为结直肠癌患者和健康个体的qPCR分析寻找最佳的参考miRNAs。485个血源性miRNAs在结直肠癌和健康对照的血清样本池中进行了分析。在30名结直肠癌患者和30名健康人中，用qPCR方法验证了从分析结果中选择的7个候选miRNAs和以前报道的3个参考miRNAs，然后用统计算法BestKeeper、geNorm和NormFinder进行了分析。综上所述，hsa-miR-93-5p、hsa-miR-25-3p和hsa-miR-106b-5p是一组合适的参考基因。更有趣的是，从485个miRNAs中验证的三个miRNAs来自单个初级转录本，表明该簇在结直肠癌中可能是高度保守的。然而，在健康人和非小细胞肺癌或乳腺癌患者之间，这三个miRNAs都有显著差异，不能作为这两种类型癌症的参考基因。
Quantitative real-time PCR (qPCR) is the most frequently used method for measuring expression levels of microRNAs (miRNAs), which is based on normalization to endogenous references. Although circulating miRNAs have been regarded as potential non-invasive biomarker of disease, no study has been performed so far on reference miRNAs for normalization in colorectal cancer. In this study we tried to identify optimal reference miRNAs for qPCR analysis across colorectal cancer patients and healthy individuals. 485 blood-derived miRNAs were profiled in serum sample pools of both colorectal cancer and healthy control. Seven candidate miRNAs chosen from profiling results as well as three previous reported reference miRNAs were validated using qPCR in 30 colorectal cancer patients and 30 healthy individuals, and thereafter analyzed by statistical algorithms BestKeeper, geNorm and NormFinder. Taken together, hsa-miR-93-5p, hsa-miR-25-3p and hsa-miR-106b-5p were recommended as a set of suitable reference genes. More interestingly, the three miRNAs validated from 485 miRNAs are derived from a single primary transcript, indicting the cluster may be highly conserved in colorectal cancer. However, all three miRNAs differed significantly between healthy individuals and non-small cell lung cancer or breast cancer patients and could not be used as reference genes in the two types of cancer.
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影响因子：
20.3
作者：
Diakos, Christofer;Zhong, Sheng;Wiemels, Joseph L.
通讯作者：
Wiemels, Joseph L.
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通讯作者：
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