A Combination of Let-7d, Let-7g and Let-7i Serves as a Stable Reference for Normalization of Serum microRNAs

被引：57

作者：

Chen, Xi ^{[1
]}

Liang, Hongwei ^{[1
]}

Guan, Danping ^{[2
]}

Wang, Cheng ^{[1
]}

Hu, Xiaoyun ^{[2
]}

Cui, Lin ^{[2
]}

Chen, Sidi ^{[3
]}

Zhang, Chunni ^{[4
]}

Zhang, Junfeng ^{[1
]}

Zen, Ke ^{[1
]}

Zhang, Chen-Yu ^{[1
]}

机构：

[1] Nanjing Univ, Sch Life Sci, Jiangsu Engn Res Ctr microRNA Biol & Biotechnol, State Key Lab Pharmaceut Biotechnol, Nanjing 210008, Jiangsu, Peoples R China

[2] Nanjing MicroMedMark Biotech Co Ltd, Nanjing, Jiangsu, Peoples R China

[3] Beijing MicroMedMark Biotech Co Ltd, Beijing, Peoples R China

[4] Nanjing Univ, Jinling Hosp, Clin Sch Med Coll, Dept Clin Lab, Nanjing, Jiangsu, Peoples R China

来源：

PLOS ONE | 2013年 / 8卷 / 11期

基金：

中国国家自然科学基金;

关键词：

CIRCULATING MICRORNAS; CANCER; EXPRESSION; BIOMARKERS; IDENTIFICATION; DIAGNOSIS; PLASMA; TARGETS;

D O I：

10.1371/journal.pone.0079652

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Recent studies have indicated that circulating microRNAs (miRNAs) in serum and plasma are stable and can serve as biomarkers of many human diseases. Measurement of circulating miRNAs with sufficient sensitivity and precision, however, faces some special challenges, among which proper normalization is the most critical but often an underappreciated issue. The primary aim of this study was to identify endogenous reference genes that maintain consistent levels under various conditions to serve as an internal control for quantification of serum miRNAs. We developed a strategy combining Illumina's sequencing by synthesis (SBS) technology, reverse transcription quantitative polymerase chain reaction (RT-qPCR) assay, literature screening and statistical analysis to screen and validate the most suitable reference genes. A combination of let-7d, let-7g and let-7i is selected as a reference for the normalization of serum miRNAs and it is statistically superior to the commonly used reference genes U6, RNU44, RNU48 and miR-16. This has important implications for proper experimental design and accurate data interpretation.

引用

页数：12

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