DNA Methylation profiles as predictors of recurrence in non muscle invasive bladder cancer: an MS-MLPA approach

Abstract

BackgroundAlthough non muscle invasive bladder cancer (NMIBC) generally has a good long-term prognosis, up to 80% of patients will nevertheless experience local recurrence after the primary tumor resection. The search for markers capable of accurately identifying patients at high risk of recurrence is ongoing. We retrospectively evaluated the methylation status of a panel of 24 tumor suppressor genes (TIMP3, APC, CDKN2A, MLH1, ATM, RARB, CDKN2B, HIC1, CHFR, BRCA1, CASP8, CDKN1B, PTEN, BRCA2, CD44, RASSF1, DAPK1, FHIT, VHL, ESR1, TP73, IGSF4, GSTP1 and CDH13) in primary lesions to obtain information about their role in predicting local recurrence in NMIBC.MethodsFormaldehyde-fixed paraffin-embedded (FFPE) samples from 74 patients operated on for bladder cancer were analyzed by methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA): 36 patients had relapsed and 38 were disease-free at the 5-year follow up. Methylation status was considered as a dichotomous variable and genes showing methylation ≥20% were defined as “positive”.ResultsMethylation frequencies were higher in non recurring than recurring tumors. A statistically significant difference was observed for HIC1 (P = 0.03), GSTP1 (P = 0.02) and RASSF1 (P = 0.03). The combination of the three genes showed 78% sensitivity and 66% specificity in identifying recurrent patients, with an overall accuracy of 72%.ConclusionsOur preliminary data suggest a potential role of HIC1, GSTP1 and RASSF1 in predicting local recurrence in NMIBC. Such information could help clinicians to identify patients at high risk of recurrence who require close monitoring during follow up.

Predictive value of gene methylation for second recurrence following surgical treatment of first bladder recurrence of a primary upper‑tract urothelial carcinoma

“…Beside its involvement in normal development in human beings, DNA methylation is frequently implicated in the onset or course of cancer due to its roles in many other regulatory processes. Several studies have reported that aberrant promoter methylation at several gene loci was associated with bladder urothelial carcinoma (13,(25)(26)(27). As bladder urothelial carcinoma and UTUC display genomic and clinical similarities, we selected 10 genes (ABCC6, BRCA1, CDH1, GDF15, HSPA2, RASSF1A, SALL3, THBS1, TMEFF2 and VIM) with a high frequency of methylation in bladder urothelial carcinoma and evaluated their methylation statuses in UTUC and their associations with clinical outcomes.…”

“…DNA extraction, bisulfite transformation and gene methylation status were evaluated according to the procedures described in our previous study (11). The methylation statuses of the genes between urothelial tumors and normal tissues were not compared as all genes investigated in this study have been validated to have a low methylation rate in normal tissues (13)(14)(15)(16).…”

Tumor Suppressor in Lung Cancer-1 Is a Prognostic Predictor for the Recurrence and Progression of Non-Muscle-Invasive Bladder Cancer

“…Interestingly, recent studies have explored the significance of gene methylation of TSLC1/IGSF4 in NMIBC samples for predicting the recurrence and molecular classification of NMIBC [28,29]. Methylation of tumor suppressor genes is known to cause the downregulation of their expression.…”

Clinical significance of protocadherin 8 (PCDH8) promoter methylation in non-muscle invasive bladder cancer

“…Thus, new biomarkers are needed to predict the outcome of bladder cancer, in addition to commonly used clinicopathological parameters [2]. In recent years, more and more researchers are interested in the aberrant methylation of different genes in bladder cancer for some reasons [9,10,26]. Firstly, aberrant methylation in the promoter regions of the tumor suppressor genes at CPG islands has been recognized as one of the hallmarks of human cancers and associated with silence of gene expression, which may be used as potential biomarker in human cancers [27][28][29][30][31].…”