Cell-free microRNA expression signatures in urine serve as novel noninvasive biomarkers for diagnosis and recurrence prediction of bladder cancer

Abstract

Urinary microRNAs (miRNAs) are potential biomarkers for the noninvasive diagnosis of bladder cancer (BC). In this study, we aimed to develop a urinary miRNAs panel for diagnosing and predicting recurrence of BC. Genome-wide miRNAs analysis by deep sequencing followed by two phases of quantitative real-time PCR assays were performed on urine supernatant of 276 BC patients and 276 controls. We identified a seven-miRNA panel (miR-7-5p, miR-22-3p, miR-29a-3p, miR-126-5p, miR-200a-3p, miR-375, and miR-423-5p) that provided high diagnostic accuracy of BC with an AUC of 0.923 and 0.916 in training and validation set, respectively. The corresponding AUCs of this panel for Ta, T1 and T2-T4 were 0.864, 0.930 and 0.978, significantly higher than those of urine cytology, which were 0.531, 0.628 and 0.724, respectively (all p < 0.05). Moreover, Kaplan–Meier analysis showed that nonmuscle-invasive BC (NMIBC) patients with high miR-22-3p and low miR-200a-3p level had worse recurrence-free survival (RFS) (p = 0.002 and 0.040, respectively). Multivariate Cox regression analysis revealed that miR-22-3p and miR-200a-3p were independently associated with RFS of NMIBC (p = 0.024 and 0.008, respectively). In conclusion, our results suggested that urinary miRNAs may have considerable clinical value in diagnosis and recurrence prediction of BC.

Circulating miRNA panels for specific and early detection in bladder cancer

“…Recently, we demonstrated the diagnostic potential of circulating miRNA in body fluids, such as breast cancer, ovarian cancer and bladder cancer . Several other research groups have performed large‐scale randomized controlled trials and confirmed the diagnostic value in independent research cohorts of bladder cancer …”

“…Recently, the molecular characterization of cell‐free DNA, circulating tumor DNA and circulating miRNA by liquid biopsy is an attractive alternative to tissue biopsy to diagnose cancer and other diseases . For diagnosis of bladder cancer by liquid biopsy, there are several reports of biomarker searches with large samples, including those detecting tumor‐specific hotspot mutations in serum DNA or miRNA in urine from patients .…”

“…For diagnosis of bladder cancer by liquid biopsy, there are several reports of biomarker searches with large samples, including those detecting tumor‐specific hotspot mutations in serum DNA or miRNA in urine from patients . For the study of serum miRNA, a few studies have investigated the potential of miRNA detection in serum for biomarkers in bladder cancer . Most of these previous reports have performed microarray or next‐generation sequencing with serum miRNA samples from patients diagnosed with bladder cancer and control individuals (non‐cancer controls) to detect preclinical bladder cancer.…”

“…19 Notably, the 7-miRNA panel in bladder cancer is also effective for the detection of both low and high grades (92.7% and 96.1% in sensitivity, respectively), Recently, the molecular characterization of cell-free DNA, circulating tumor DNA and circulating miRNA by liquid biopsy is an attractive alternative to tissue biopsy to diagnose cancer and other diseases. [13][14][15][20][21][22][23] In addition, urinary cytology, which has high specificity but low sensitivity, is commonly used for detecting bladder cancer. In the bladder cancer group, many cases also had not been captured by urinary cytology (Table 1).…”

“…[10][11][12] Several other research groups have performed large-scale randomized controlled trials and confirmed the diagnostic value in independent research cohorts of bladder cancer. 13,14 Here, we analyzed 972 serum samples, including 392 bladder cancer samples, 100 non-cancer control samples and 480 samples of other types of tumors, by performing a comprehensive miRNA microarray. In the training set, we identified 7 miRNA candidates for bladder cancer screening.…”