A direct quantification method for measuring plasma MicroRNAs identified potential biomarkers for detecting metastatic breast cancer
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Qian Zhao1,*, Shengqiong Deng1,*, Guangxue Wang1,*, Cuicui Liu2, Lingyu Meng2, Shanshan Qiao1, Lei Shen1, Yue Zhang1, Jinhui Lü3, Wenshu Li3, Yuzhen Zhang1, Min Wang4, Richard G. Pestell4, Chunli Liang1, Zuoren Yu1,2,3
1Research Center for Translational Medicine, Translational Medical Center for Stem Cell Therapy, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China
2Shanghai East Hospital, Dalian Medical University, Dalian, China
3School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou, China
4Department of Cancer Biology, Sidney Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, USA
*These authors contributed equally to this work
Zuoren Yu, e-mail: [email protected]
Keywords: circulating miRNA, direct quantification, biomarker, metastatic breast cancer, miR-106a
Received: November 29, 2015 Accepted: February 21, 2016 Published: March 08, 2016
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.
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