Identification of a three-miRNA panel in serum for bladder cancer diagnosis by a diagnostic test

Abstract

Background Bladder cancer (BC) is the tenth most common cancer in the world. Serum microRNA (miRNA) profiles previously have been reported as non-invasive biomarkers in cancer screening. The non-invasive and reliable diagnostic biomarkers are urgently needed for detecting BC, while cystoscopy is invasive. Our study aimed to identify candidate miRNAs in serum as potential diagnostic biomarkers for BC detection. Methods This study was including the screening stage, training stage, and validation stage with 137 BC patients and 127 healthy controls (HCs). We identified the expression of 28 serum miRNAs from 5 BC pools and 3 HC pools in the initial screening stage. The other 112 BC patients and 112 HCs were randomly divided into training stage with 30 BC patients and 30 HCs and validation stages with 82 BC patients and 82 HCs. These HCs matched BC patients based on age and gender with P value >0.05. Identified dysregulated miRNAs were further confirmed in the training stage, and validation stages by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The diagnostic value of miRNAs was assessed by receiver operating characteristic (ROC) curves and the area under the ROC curve (AUC). Target genes of 3 candidate miRNAs were predicted by bioinformatic analysis. Results Five miRNAs (miR-106a-5p, miR-145-5p, miR-132-3p, miR-7-5p and miR-148b-3p) in serum were obviously dysregulated in BC patients compared to HCs. The ability to diagnose BC of 3 candidate miRNAs was estimated by AUC, with miR-132-3p (AUC =0.781; sensitivity =68.29%, specificity =81.71%), miR-7-5p (AUC =0.778; sensitivity =59.76%, specificity =84.15%) and miR-148b-3p (AUC =0.837; sensitivity =81.71%, specificity =71.95%). Combined application of these candidate miRNAs with parallel test could improve the diagnostic value (AUC =0.922; sensitivity =90.24%, specificity =81.71%). BNC2, GAS7 , and NTRK2 , considered as target genes of the three-miRNA panel, may play an important role in the process of BC development. Conclusions A three-miRNA panel in serum was identified for BC diagnosis in our study, which HCs were used for differential diagnosis. The three-miRNA panel (miR-132-3p, miR-7-5p, and miR-148b-3p) might be performed as a non-invasive and convenient diagnostic tool for BC screening and diagnosis.

Basonuclin-2 regulates extracellular matrix production and degradation

“…Coupled with observations that BNC2 expression is lower in ovarian cancer cell lines compared with normal tissue (14) and that the deletion of SNP-containing regions decreases expression (11), BNC2 was thereby implicated as an important cancer-associated gene. Similar studies also note the association of BNC2-related SNPs across a range of cancer types (esophageal (15), prostate (16), bladder (17,18,19), liver (20), skin (21,22,23), uterus (24,25), stomach (26), and lung (27)).…”

Maternal hsa-miR-423-5p associated with the cognitive development of babies in pregnant women without mental disorders

“…At this time, as an initial precaution, we quantified the samples using the Spectrophotometer (Thermo Scientific™ NanoDrop™) to evaluate the purification of the extracted RNA. Regarding endogenous control and duplicate reactions, we did not find standardization in the literature, including studies published in recent years using a single endogenous miRNA for control ( Sekovanić et al, 2021 ; Bayati et al, 2022 ; Couto et al, 2022 ; Li et al, 2022 ; Ding et al, 2023 ; Higashi et al, 2023 ). Furthermore, Couto et al (2022) cite duplicate reactions and the other authors do not cite the number of reactions carried out.…”

Potential clinical application of microRNAs in bladder cancer

“…Higher expression of miR-152 as well as lower expression of both miR-3187-3p and miR-27a-3p are associated with characteristics of a poorer prognosis of BC patients with characteristics of advanced clinical stage, higher tumor grade, and positive lymph node metastasis. Specifically, NMIBC patients with high miR-152 expression and low miR-3187-3p expression had a poor relapse-free survival, miR-152 was independently associated with tumor recurrence in NMIBC [41] , and miR-20a-5p was also associated with BC recurrence [40] . In addition, the expression levels of miR-210 were associated with the staging and grading of BC, and the expression of serum miR-210 was significantly decreased in postoperative paired specimens, but increased in most patients with recurrent BC [45] (Table 1).…”

“…In contrast to microarray screening, however, high-throughput sequencing eliminates some potential contamination of other small RNA and DNA fragments. For example, Jiang et al [41] employed MiSeq to identify 26 differentially expressed miRNAs in BC, and found that the AUC for the combination of six miRNAs used for early BC diagnosis in patients with Ta and T1 stages was significantly higher (0.841), compared with that of urine cytology (0.645) [41] . In a study conducted by Li et al, a combination of four miRNAs was identified for BC diagnosis in the serum, demonstrating an exceptional diagnostic power with an AUC of 0.985 [42] .…”