Zhan Cheng, Yan Li, Wang Lin, Jiang Wei, Zhang Yongxing, Xi Junjie, Chen Li, Jin Yulin, Qiao Yulei, Shi Yu, Wang Qun
Department of Thoracic Surgery, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
Department of Radiation Oncology, Eye and ENT Hospital, Fudan University, Shanghai 200031, China.
Biochem Biophys Res Commun. 2014 Oct 24;453(3):375-8. doi: 10.1016/j.bbrc.2014.09.086. Epub 2014 Sep 27.
Although the accuracy of detecting the expression of miRNAs by quantitative real-time polymerase chain reaction (qRT-PCR) is highly dependent on reliable reference miRNAs, many commonly used reference miRNAs are not stably expressed and as such are not suitable for quantification and normalization of qRT-PCR data. To solve this problem, we analyzed the global expression profiles of thousands of samples in 14 types of common human tumors released by The Cancer Genome Atlas (TCGA), and identified the most stably and highly expressed miRNAs as candidate reference miRNAs in each type of tumor. We found that miR-361-5p and let-7i-5p were the most recommended candidate reference miRNAs in nine and eight types of tumors, respectively, followed by let-7a-5p, mir-28-5p and miR-99b-5p. Our results are of important value to those researchers focused on miRNA; however, these candidate reference miRNAs still need to be validated prior to their use in qRT-PCR studies.
尽管通过定量实时聚合酶链反应(qRT-PCR)检测微小RNA(miRNA)表达的准确性高度依赖于可靠的内参miRNA,但许多常用的内参miRNA表达并不稳定,因此不适用于qRT-PCR数据的定量和标准化。为了解决这个问题,我们分析了癌症基因组图谱(TCGA)发布的14种常见人类肿瘤中数千个样本的整体表达谱,并确定了每种肿瘤中最稳定且高表达的miRNA作为候选内参miRNA。我们发现,miR-361-5p和let-7i-5p分别是九种和八种肿瘤中最推荐的候选内参miRNA,其次是let-7a-5p、mir-28-5p和miR-99b-5p。我们的结果对专注于miRNA的研究人员具有重要价值;然而,这些候选内参miRNA在用于qRT-PCR研究之前仍需验证。