A Panel of Urinary Long Non-coding RNAs Differentiate Bladder Cancer from Urocystitis

Abstract

Liquid biopsy is becoming a promising method for non-invasive cancer detection. In several proof-of-concept studies, long non-coding RNAs (lncRNAs) were found to be potential biomarkers for bladder cancer detection. The objective of this study was to discover a panel of cell-free, urinary lncRNAs as liquid biopsy biomarkers to non-invasively differentiate bladder cancer from chronic urocystitis. To this end, we collected urine samples from both bladder cancer patients and urocystitis patients. These samples were divided into discovery group and validation group. In the discovery group, the expression levels of 16 cell-free urinary lncRNAs were measured by qPCR to discover candidate biomarkers. The diagnostic performance of the candidate lncRNA biomarkers was then evaluated, which led to a panel of lncRNA biomarkers for bladder cancer detection. The performance of this panel of biomarkers was further evaluated in the validation group to see if these lncRNA biomarkers could discriminate the bladder cancer patients from urocystitis patients. We found that all of the 16 lncRNAs evaluated in this study demonstrated significant difference (p<0.05) of expression between bladder cancer patients and urocystitis patients. Nine lncRNAs provided decent diagnostic performance with area under the receiver operating characteristic (ROC) curve (AUC) reaching 0.70 or higher. We then selected the top four lncRNAs, namely UCA1-201, HOTAIR, HYMA1 and MALAT1, to form a panel of urinary biomarkers. Using this panel, bladder cancer patients could be discriminated from urocystitis patients, with sensitivity and specificity reaching 95.7% and 94.3%, respectively. Finally, we confirmed the applicability of the four-lncRNA panel in an independent validation study that included 60 bladder cancer patients and 60 urocystitis patients. Our study paves the way for further studies aimed at large-scale clinical tests of developing lncRNA biomarkers in urine for bladder cancer diagnostics.

Applying Serum Proteins and MicroRNA as Novel Biomarkers for Early-Stage Cervical Cancer Detection

“…The cycling parameters for miRNA were as follows: 95 °C for 30 sec, followed by 40 cycles of 95 °C for 5 sec and 60 °C for 30 sec. Finally, we quantified the miRNA level by using the ΔCt method that was previously described [37][38][39] . In short, the miRNA level = 2^ ΔCt multiplying 1000, where ΔCt was the difference between Ct value of the miRNA of interest to Ct value of internal controls.…”

Regulatory role of long non‑coding RNA UCA1 in signaling pathways and its clinical applications (Review)

“…Early differential diagnosis of cancer and precancerous lesions is an important method to decrease the risk of recurrence and improve prognosis (121). A study has demonstrated that the combination of UCA1, HOX transcript antisense intergenic RNA, hydatidiform mole associated and imprinted and metastasis-associated lung adenocarcinoma transcript 1 can distinguish between patients with bladder cancer and patients with urocystitis, with a sensitivity and specificity of 95.7% and 94.3%, respectively (121). Collectively, these findings suggest that it is feasible to use UCA1 as a diagnostic biomarker.…”

Tumor heterogeneity and the potential role of liquid biopsy in bladder cancer

“…The results are very heterogeneous, but promising candidate nucleic acids biomarkers have been identified with higher sensitivities and specificities than plasma ctDNA. The major studies that have investigated the role of exosomes in the diagnosis of BC are compiled in Table 3 [86,[97][98][99][100][101][102][103][104]. Although the specific functional role of exosomes in BC heterogeneity remains unclear, exosomes have emerged as potential noninvasive disease biomarkers with potential clinical significance.…”

“…An alternative strategy to understand tumor heterogeneity might be liquid biopsy, which can detect genetic alterations and tumor cells present in peripheral blood or urine of patients with BC [105,99]. Peripheral blood-or urine-based genomic analyses were flexible in processing, and there are no specific device required for isolation, which have the potential to diagnose and dynamically monitor BC recurrence [82,78,106,69,65,79].…”