Document Type
Article
Publication Date
4-19-2016
Abstract
Circulating miRNAs are protected from ribonuclease degradation by assembly into microvesicles and exosomes. Releasing miRNAs completely from these particles is the key step to quantify the circulating miRNAs. Currently purified RNA-based quantitative analysis is widely used while it is time and cost consuming with high risk for those circulating miRNAs with low abundance due to partial loss of RNA during the steps of total RNA extraction and small RNA enrichment. Herein, we optimized a simple, effective and time-saving method to directly measure plasma miRNAs without RNA isolation. It is based on complete miRNA release from the protein complexes, followed by miRNA-specific reverse transcription and quantitative real-time PCR amplification. By comparison to the RNA-based approach, the direct quantification method showed more efficiency for circulating miRNA analysis, higher accuracy and specificity. By application of the direct quantification method to clinical samples combined with the RNA-based miRNA screening analysis, upregulation of miR-106a in blood was validated in metastatic breast cancer patients, indicating miR-106a are a potential biomarker for metastatic breast cancer.
Recommended Citation
Zhao, Qian; Deng, Shengqiong; Wang, Guangxue; Liu, Cuicui; Meng, Lingyu; Qiao, Shanshan; Shen, Lei; Zhang, Yue; Lü, Jinhui; Zhang, Yuzhen; Wang, Min; Pestell, Richard; Liang, Chunli; Yu, Zuoren; and Li, Wenshu, "A direct quantification method for measuring plasma MicroRNAs identified potential biomarkers for detecting metastatic breast cancer." (2016). Department of Cancer Biology Faculty Papers. Paper 86.
https://jdc.jefferson.edu/cbfp/86
Creative Commons License
This work is licensed under a Creative Commons Attribution 3.0 License.
PubMed ID
26967564
Comments
This article has been peer reviewed. It was published in: Oncotarget.
Volume 7, Issue 16, 19 April 2016, Pages 21865-21874.
The published version is available at DOI: 10.18632/oncotarget.7990
Copyright © 2016 The Authors