title: Microfluidic-based multiplex qRT-PCR identifies diagnostic and prognostic microRNA signatures in the sera of prostate cancer patients.
abstract: Recent prostate-specific antigen-based screening trials indicate an urgent need for novel and noninvasive biomarker identification strategies to improve the prediction of prostate cancer behavior. Noncoding microRNAs (miRNA) in the serum and plasma have been shown to have potential as noninvasive markers for physiologic and pathologic conditions. To identify serum miRNAs that diagnose and correlate with the prognosis of prostate cancer, we developed a multiplex quantitative reverse transcription PCR method involving the purification of multiplex PCR products followed by uniplex analysis on a microfluidics chip to evaluate 384 human miRNAs. Using Dgcr8 and Dicer knockout (small RNA-deficient) mouse ES cells as the benchmark, we confirmed the validity of our technique and uncovered a considerable lack of accuracy in previously published methods. Profiling 48 sera from healthy men and untreated prostate cancer patients with differing CAPRA scores, we identified miRNA signatures that allow us to diagnose cancer patients and correlate with a prognosis. These serum signatures include oncogenic and tumor-suppressive miRNAs, suggesting functional roles in prostate cancer progression.
authors.affiliation: The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Department of Urology, UCSF-Helen Diller Family Comprehensive Cancer Center, University of California San Francisco, San Francisco, California 94143, USA.
authors.forename: Felix
authors.initials: F
authors.lastname: Moltzahn
authors.forename: Adam B
authors.initials: AB
authors.lastname: Olshen
authors.forename: Lauren
authors.initials: L
authors.lastname: Baehner
authors.forename: Andrew
authors.initials: A
authors.lastname: Peek
authors.forename: Lawrence
authors.initials: L
authors.lastname: Fong
authors.forename: Hubert
authors.initials: H
authors.lastname: Stöppler
authors.forename: Jeffry
authors.initials: J
authors.lastname: Simko
authors.forename: Joan F
authors.initials: JF
authors.lastname: Hilton
authors.forename: Peter
authors.initials: P
authors.lastname: Carroll
authors.forename: Robert
authors.initials: R
authors.lastname: Blelloch
mesh_terms: D016022:Case-Control Studies; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000097:blood* / Q000235:genetics; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D011379:Prognosis; D011471:Prostatic Neoplasms / Q000097:blood* / Q000175:diagnosis / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
chemical_list: D035683:MicroRNAs