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：

070301 [无机化学]; 070403 [天体物理学]; 070507 [自然资源与国土空间规划学]; 090105 [作物生产系统与生态工程];

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

共 41 条

[1]

Circulating microRNA-1 as a potential novel biomarker for acute myocardial infarction [J].

Ai, Jing ;

Zhang, Rong ;

Li, Yue ;

Pu, Jielin ;

Lu, Yanjie ;

Jiao, Jundong ;

Li, Kang ;

Yu, Bo ;

Li, Zhuqin ;

Wang, Rongrong ;

Wang, Lihong ;

Li, Qiang ;

Wang, Nina ;

Shan, Hongli ;

Li, Zhongyu ;

Yang, Baofeng .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2010, 391 (01) :73-77

[2]

The functions of animal microRNAs [J].

Ambros, V .

NATURE, 2004, 431 (7006) :350-355

[3]

Normalization of real-time quantitative reverse transcription-PCR data: A model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets [J].

Andersen, CL ;

Jensen, JL ;

Orntoft, TF .

CANCER RESEARCH, 2004, 64 (15) :5245-5250

[4]

MicroRNA expression profiles predictive of human renal allograft status [J].

Anglicheau, Dany ;

Sharma, Vijay K. ;

Ding, Ruchuang ;

Hummel, Aurelie ;

Snopkowski, Catherine ;

Dadhania, Darshana ;

Seshan, Surya V. ;

Suthanthiran, Manikkam .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2009, 106 (13) :5330-5335

[5]

Argonaute2 complexes carry a population of circulating microRNAs independent of vesicles in human plasma [J].

Arroyo, Jason D. ;

Chevillet, John R. ;

Kroh, Evan M. ;

Ruf, Ingrid K. ;

Pritchard, Colin C. ;

Gibson, Donald F. ;

Mitchell, Patrick S. ;

Bennett, Christopher F. ;

Pogosova-Agadjanyan, Era L. ;

Stirewalt, Derek L. ;

Tait, Jonathan F. ;

Tewari, Muneesh .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2011, 108 (12) :5003-5008

[6]

MicroRNAs: Genomics, biogenesis, mechanism, and function (Reprinted from Cell, vol 116, pg 281-297, 2004) [J].

Bartel, David P. .

CELL, 2007, 131 (04) :11-29

[7]

Characterization of microRNAs in serum: a novel class of biomarkers for diagnosis of cancer and other diseases [J].

Chen, Xi ;

Ba, Yi ;

Ma, Lijia ;

Cai, Xing ;

Yin, Yuan ;

Wang, Kehui ;

Guo, Jigang ;

Zhang, Yujing ;

Chen, Jiangning ;

Guo, Xing ;

Li, Qibin ;

Li, Xiaoying ;

Wang, Wenjing ;

Zhang, Yan ;

Wang, Jin ;

Jiang, Xueyuan ;

Xiang, Yang ;

Xu, Chen ;

Zheng, Pingping ;

Zhang, Juanbin ;

Li, Ruiqiang ;

Zhang, Hongjie ;

Shang, Xiaobin ;

Gong, Ting ;

Ning, Guang ;

Wang, Jun ;

Zen, Ke ;

Zhang, Junfeng ;

Zhang, Chen-Yu .

CELL RESEARCH, 2008, 18 (10) :997-1006

[8]

Identification of ten serum microRNAs from a genome-wide serum microRNA expression profile as novel noninvasive biomarkers for nonsmall cell lung cancer diagnosis [J].

Chen, Xi ;

Hu, Zhibin ;

Wang, Wenjing ;

Ba, Yi ;

Ma, Lijia ;

Zhang, Chunni ;

Wang, Cheng ;

Ren, Zhiji ;

Zhao, Yang ;

Wu, Sijia ;

Zhuang, Rui ;

Zhang, Yixin ;

Hu, Heng ;

Liu, Chazhen ;

Xu, Lin ;

Wang, Jun ;

Shen, Hongbing ;

Zhang, Junfeng ;

Zen, Ke ;

Zhang, Chen-Yu .

INTERNATIONAL JOURNAL OF CANCER, 2012, 130 (07) :1620-1628

[9]

Identification and characterization of microRNAs in raw milk during different periods of lactation, commercial fluid, and powdered milk products [J].

Chen, Xi ;

Gao, Chao ;

Li, Haijin ;

Huang, Lei ;

Sun, Qi ;

Dong, Yanye ;

Tian, Chunliang ;

Gao, Shengpu ;

Dong, Hailin ;

Guan, Danping ;

Hu, Xiaoyun ;

Zhao, Shujian ;

Li, Liang ;

Zhu, Lin ;

Tan, Qiao ;

Zhang, Junfeng ;

Zen, Ke ;

Zhang, Chen-Yu .

CELL RESEARCH, 2010, 20 (10) :1128-1137

[10]

MicroRNA expression profiling during human cord blood-derived CD34 cell erythropoiesis [J].

Choong, Meng Ling ;

Yang, Henry He ;

McNiece, Ian .

EXPERIMENTAL HEMATOLOGY, 2007, 35 (04) :551-564

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