Serum miRNA profiling and validation.

<p>(<i>A)</i> Measurement of circulating miRNAs in sera pooled from patients with advanced prostate cancer as compared to healthy donors (comprising a Discovery Set) by TLDA profiling. Blue- and brown-filled circles represent serum miRNAs increased or decreased (with unadjusted p-value <0.05), respectively, in mCRPC patients compared to healthy controls. <i>Inset:</i> Nine miRNAs demonstrated >5-fold change (unadjusted <i>P</i><0.05, Student’s t-test). FC, fold-change. (<i>B)</i> Confirmation of mCRPC-associated serum miRNAs in individual samples from the Discovery Set from the University of Washington samples. <i>Upper</i>: miRNA biomarker candidates were measured in individual samples by TaqMan miRNA qRT-PCR (<i>P</i> value assigned by Wilcoxon signed-rank test), where miRNA abundance is given in terms of miRNA copies/µl serum. Red bars, mean +/− SEM of miRNA copies/µl serum for each group. <i>Lower</i>: Receiver operating characteristic (ROC) curves plot sensitivity vs. (1 - specificity) to assess the ability of each miRNA biomarker to distinguish cases from controls. <i>(C)</i> Validation of mCRPC-associated serum miRNAs in an independent Validation Set. <i>Upper</i>: Serum concentration (copies/µl) of miR-141, miR-375, miR-200c, miR-200a and miR-210 was measured by TaqMan miRNA qRT-PCR. Dot-plot associated <i>P</i> values were assigned by Wilcoxon signed-rank test. Dot plots and ROC curves were generated as described for <b>Fig. 1</b>. <i>Lower</i>: <i>Red</i>, results from the validation sample set obtained from the University of Michigan. <i>Black</i>, results from the primary sample set obtained from the University of Washington reproduced from <b>Fig. 1</b><b><i>B</i></b><i>, lower</i>. AUC, area under the curve; mCRPC, prostate cancer patient sera; FC, fold-change; CTL, control sera (from age-matched male individuals with normal PSA and negative digital rectal exam).