Improved Noninvasive Bladder Cancer Diagnosis using Urine Sediments and Novel DNA Methylation Biomarker Panels

Background: Aberrant DNA methylation status early in carcinogenesis represents a potential indicator of tumor detection. We would like to establish a DNA methylation biomarker panel for bladder cancer detection in the research. Methods: Seven candidate genes with known cancer associations were selected for this study. The DNA methylation status of the candidate genes was analyzed by methylation-specific polymerase chain reaction assays to evaluate the relationship between bladder cancer and target gene methylation status in the urine sediments of participants. Results: 112 bladder cancer patients, 10 healthy volunteers, and 17 glandular cystitis patients were enrolled. There were significant differences in the methylation status of p14ARF, RUNX3, RAR beta, DAPK, and HPP1 between the healthy control, glandular cystitis, and bladder cancer groups (p = 0.027, p < 0.001, p < 0.001, p = 0.030 and p = 0.003, respectively). A panel composed of all five genes with significant methylation differences yielded an area under the receiver-operating characteristic curve (AUC) of 0.936 and had 98.21% sensitivity and 88.89% specificity, while a panel using just two of these genes (RUNX3 and RAW yielded an AUC of 0.918 with 96.64% sensitivity and 88.89% specificity. Another panel of two genes (p14ARF and HPP1) had 100% specificity, but an AUC of 0.688 and 37.50% sensitivity. Conclusions: Using the urine of bladder cancer patients and healthy controls, we assessed several novel DNA methylation biomarker panels that demonstrated good bladder cancer detection capability. Based on the results of this study we recommend the RUNX3 and RARE panel as the first choice for bladder cancer detection. In suspicious or difficult to diagnose cases, the p14ARF and HPP1 panel could be used as an additional diagnostic tool due to its high specificity.