Guo Yuanyuan, Zhou Xuantong, Gao Feng, Wang Minjie, Yang Qi, Li Xin, Liu Zhihua, Luo Aiping
The State Key Laboratory of Molecular Oncology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100021, China.
Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Department of Thoracic Surgery II, Peking University Cancer Hospital & Institute, Beijing 100142, China.
Heliyon. 2023 Mar 25;9(4):e14515. doi: 10.1016/j.heliyon.2023.e14515. eCollection 2023 Apr.
Circulating miRNA expression is most commonly measured by qRT-PCR, however, the lack of a suitable endogenous control hinders people from evaluating the accurate changes in miRNA expression levels and developing the non-invasive biomarkers. In this study, we aimed to screen the specific, highly stable endogenous control in esophageal squamous cell carcinoma (ESCC) to overcome the obstacle. We selected "housekeeping" miRNAs according to the published database and initially acquired 21 miRNAs. Subsequently, we screened these miRNAs using GSE106817 and TCGA datasets according to specific inclusion criteria and evaluated the suitability of "candidate" miRNAs. Among these miRNAs, the average abundance of miR-423-5p was relatively high in serum. Notably, miR-423-5p expression in serum showed no significant difference between ESCC patients and healthy controls (n = 188, P = 0.29). Moreover, among these miRNAs, miR-423-5p was the most stable miRNA using the NormFinder algorithms. Overall, these results indicate that miR-423-5p, as a novel and optimal endogenous control, could be used to quantify circulating miRNAs in ESCC.
循环miRNA的表达最常通过qRT-PCR进行检测,然而,缺乏合适的内参阻碍了人们评估miRNA表达水平的准确变化以及开发非侵入性生物标志物。在本研究中,我们旨在筛选食管鳞状细胞癌(ESCC)中特异性、高度稳定的内参,以克服这一障碍。我们根据已发表的数据库选择了“管家”miRNA,最初获得了21种miRNA。随后,我们根据特定的纳入标准,使用GSE106817和TCGA数据集对这些miRNA进行筛选,并评估“候选”miRNA的适用性。在这些miRNA中,血清中miR-423-5p的平均丰度相对较高。值得注意的是,ESCC患者与健康对照者血清中miR-423-5p的表达无显著差异(n = 188,P = 0.29)。此外,在这些miRNA中,使用NormFinder算法分析发现miR-423-5p是最稳定的miRNA。总体而言,这些结果表明,miR-423-5p作为一种新型的最佳内参,可用于定量ESCC中循环miRNA。