Aberrant Methylation ofRARβ2andAPCGenes in Voided Urine as Molecular Markers for Early Detection of Bilharzial and Nonbilharzial Bladder Cancer

Abstract

Background: Bladder cancer cells illustrate major disruptions in their DNA methylation patterns as compared with normal ones. Authors aimed to identify epigenetic molecular markers in urine for early detection of bladder cancer.Materials and Methods: We retrospectively analyzed the methylation status of RARb 2 and APC genes in urine samples from 210 bladder cancer patients, 61 patients with benign urological diseases, and 49 healthy volunteers by using methylation-specific PCR.Results: Methylated RARb 2 and APC were significantly higher in bladder cancer patients (62.8%, 59.5%) than benign (16.4%, 5%) but not detected in healthy volunteers (0%) at (P < 0.0001). Both methylated genes showed no significant difference among clinicopathologic factors; however, they were detected in all grades and stages. Among the 128 patients with bilharzial bladder cancer, 94 (73.4%) showed methylated RARb 2 and 86 (67.2%) showed methylated APC. Homoplasmic methylation pattern of both genes were only detected in bilharzial bladder cancer cases. Both sensitivities and specificities of the methylated genes for bladder cancer detection were superior to urine cytology and when altogether combined, the sensitivities improved to (91.8%), (93.5%), (91.9%), and (80.9%) in detection of: bladder cancer, non-muscle invasive bladder cancer, low-grade tumors, and bilharzial associated bladder cancer, respectively.Conclusion: Thus, methylated RARb 2 and APC genes might be valuable urinary molecular markers for early detection of bilharzial and nonbilharzial bladder cancer. Cancer Epidemiol Biomarkers Prev; 20(8); 1657-64. Ó2011 AACR.

Molecular markers and Schistosoma-associated bladder carcinoma: A systematic review and meta-analysis

“…However, there was potential publication bias in the quantitative analysis of p16 deletion and p27. The association of telomerase by TERT [31][32][33]57] or TRAP [31][32][33]58,59], sFas [8,32,34], CD44 [37,[60][61][62], and RARb2 [46,63,64] with schistosomal bladder cancer is shown in Fig. 4.…”

Urine biomarkers of schistosomiais and its associated bladder cancer

“…Homoplasmic methylation pattern of both genes was only detected in schistosomal BC cases. Both sensitivities and specificities of the methylated genes for BC detection were superior to urine cytology and when altogether combined, the sensitivities improved to (91.8%), (93.5%), (91.9%) and (80.9%) in detection of BC, non-muscle invasive BC, lowgrade tumors and schistosomal-associated BC, respectively [61]. A. Biomarkers related to cell cycle…”

Hypermethylation in bladder cancer: biological pathways and translational applications

“…These include fine analyses of genomic regions being hypermethylated [56,128], and individual tumor suppresor candidates such as the Runt-related transcription factor 3 gene (RUNX3) [100,[129][130][131][132], myopodin, a cytoskeleton actin binding protein whose methylation was found useful to predict BCG response [133][134][135], the phosphatase and tensin homolog gene (PTEN) [58][59][60]130], the hypermethylated in cancer 1 gene (HIC1) [86,87,130], the Von Hippel-Lindau tumor suppresor gene (VHL) [58,59,68], the Wilms tumor 1 gene (WT1) [59,60], the deleted in bladder cancer,chromosomal region candidate 1 gene (DBCCR1) [128], the immunoglobulin superfamily member 4 gene (IGSF4) [58,59,91], or the deleted in breast cancer 1 gene (DBC1) [140]. Differentiation related genes have also been found methylated with high prevalence in bladder tumors such as the S-100 family of proteins [136], the Sry-related-hmg box 9 gene (SOX9) [47], the polyamine modulated factor 1 gene (PMF1) [137], the cancer antigen 1 gene (CAGE1) [130,138], the estrogen receptor gene (ESR1) [58][59][60]87], or the retinoic acid receptor gene (RARB) [58,59,105,139,…”

“…Differentiation related genes have also been found methylated with high prevalence in bladder tumors such as the S-100 family of proteins [136], the Sry-related-hmg box 9 gene (SOX9) [47], the polyamine modulated factor 1 gene (PMF1) [137], the cancer antigen 1 gene (CAGE1) [130,138], the estrogen receptor gene (ESR1) [58][59][60]87], or the retinoic acid receptor gene (RARB) [58,59,105,139,140]. Some of these genes have been shown useful for characterizing specific tumor subtypes, such as differentiating Bilharzial from nonbilharzial tumors [139], or characterizing small-cell carcinomas of the bladder [141]. In this review, the focus has dealt on genes studied in high number of tumors or in several reports.…”

“…The initial studies have tried to evaluate whether urinary methylomes may serve for the diagnosis of the disease ( Table 1). The most studied genes reporting to play a potential diagnostic role with global accuracies over 65% in urinary specimens include CDKN2A [57, 58, 60, 67, 79, 83-85, 108, 115, 127], CCND2 [83], GSTP1 [58, 83-85, 108, 131], APC [57, 60, 83-85, 108, 142, 143], BRCA1 [58,131], RASFF1A [60, 71, 74, 79, 83-85, 105, 108, 115, 131, 142, 143], BCL2 [60,74,101,131,142], CDH1 [60,84,105,108,127], SFRP1 [57,115,124,127,140], SFRP2,4,5 [124,140], WIF1 [60,124,140,142], myopodin [134], PMF1 [137], and RARB [57,58,60,67,84,85,105,108,139,140], among others. These studies were initially performed by MS-PCR in qualitative or quantitative fashion modalities [67, 71, 74, 79, 83-85, 101, 105, 108, 110, 115, 119, 124, 126, 131, 139, 140, 142] and currently it is feasible by comprehensive multiplexed analyses such as by MS-MLPA …”