title: A direct quantification method for measuring plasma MicroRNAs identified potential biomarkers for detecting metastatic breast cancer.
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.
authors.affiliation: Research Center for Translational Medicine, Translational Medical Center for Stem Cell Therapy, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Zhao
authors.affiliation: Research Center for Translational Medicine, Translational Medical Center for Stem Cell Therapy, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China.
authors.forename: Shengqiong
authors.initials: S
authors.lastname: Deng
authors.affiliation: Research Center for Translational Medicine, Translational Medical Center for Stem Cell Therapy, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China.
authors.forename: Guangxue
authors.initials: G
authors.lastname: Wang
authors.affiliation: Shanghai East Hospital, Dalian Medical University, Dalian, China.
authors.forename: Cuicui
authors.initials: C
authors.lastname: Liu
authors.affiliation: Shanghai East Hospital, Dalian Medical University, Dalian, China.
authors.forename: Lingyu
authors.initials: L
authors.lastname: Meng
authors.affiliation: Research Center for Translational Medicine, Translational Medical Center for Stem Cell Therapy, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China.
authors.forename: Shanshan
authors.initials: S
authors.lastname: Qiao
authors.affiliation: Research Center for Translational Medicine, Translational Medical Center for Stem Cell Therapy, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Shen
authors.affiliation: Research Center for Translational Medicine, Translational Medical Center for Stem Cell Therapy, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou, China.
authors.forename: Jinhui
authors.initials: J
authors.lastname: Lü
authors.affiliation: School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou, China.
authors.forename: Wenshu
authors.initials: W
authors.lastname: Li
authors.affiliation: Research Center for Translational Medicine, Translational Medical Center for Stem Cell Therapy, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China.
authors.forename: Yuzhen
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Cancer Biology, Sidney Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, USA.
authors.forename: Min
authors.initials: M
authors.lastname: Wang
authors.affiliation: Department of Cancer Biology, Sidney Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, USA.
authors.forename: Richard G
authors.initials: RG
authors.lastname: Pestell
authors.affiliation: Research Center for Translational Medicine, Translational Medical Center for Stem Cell Therapy, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China.
authors.forename: Chunli
authors.initials: C
authors.lastname: Liang
authors.affiliation: Research Center for Translational Medicine, Translational Medical Center for Stem Cell Therapy, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China.|Shanghai East Hospital, Dalian Medical University, Dalian, China.|School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou, China.
authors.forename: Zuoren
authors.initials: Z
authors.lastname: Yu
mesh_terms: D014408:Biomarkers, Tumor / Q000097:blood* / Q000235:genetics; D001774:Blood Chemical Analysis / Q000379:methods*; D001943:Breast Neoplasms / Q000097:blood* / Q000235:genetics; D005260:Female; D006801:Humans; D035683:MicroRNAs / Q000097:blood*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D014408:Biomarkers, Tumor; D035683:MicroRNAs
keywords: biomarker; circulating miRNA; direct quantification; metastatic breast cancer; miR-106a