Improved detection of bladder carcinoma cells in voided urine by standardized microsatellite analysis

Abstract

Successful treatment of bladder cancer depends largely on early diagnosis of primary and recurrent disease. Sensitive, specific and noninvasive procedures for detection are especially needed for grade 1 and 2 bladder tumors, because of the relatively low sensitivity of cytology. Here we introduce a novel strategy to improve the sensitivity and reliability of microsatellite analyses by employing marker-specific threshold values for loss-of-heterozygosity (LOH) at 10 loci. These individual cut-offs were experimentally determined with 35 normal control tissues and subsequently validated in a retrospective study with bladder cancer biopsies from 86 patients. In a prospective analysis of voided urines samples and matched blood probes from 91 patients, LOH-analysis, UroVysion FISH and conventional urine cytology were compared with histological findings of consecutive transurethral biopsies. Whereas all samples could be analyzed by our LOH assay, only 56 samples were suitable for all 3 analyses. The highest sensitivity was obtained with our LOH-assay/cytology approach (G1-2: 72%; G3: 96%) being only surpassed by a combination of all 3 techniques (G1-2: 83%; G3: 100%). Since over 93% of the patients with recurrent disease were identified by LOH/cytology-analyses of their voided urine samples, a monitoring scheme alternating cystoscopy with LOH/cytology-examination could now be envisioned to reduce invasive interventions and consequently improve followup compliance, especially in patients with low grade tumors. ' 2007 Wiley-Liss, Inc.

BAMBI gene is epigenetically silenced in subset of high‐grade bladder cancer

“…1 Of bladder tumors, 70-80% are, at the time of initial examination, detected as low-grade noninvasive or minimally invasive papillary tumors (pTa or pT1) with a high rate of recurrence, but with a low tendency for progression to advanced disease. 2,3 The remaining 20% are, despite improvements in diagnostic methods and treatments, detected as high-grade invasive cancer with poor prognosis, 4 and seem as either superficial spreading flat (carcinoma in situ/pTis) or invasive nodular tumors. By conventional histopathological grading and staging of bladder cancer, however, especially at initial biopsy obtained from the superficial area or urinary cytological samples, it is often difficult to foresee which tumors will recur or progress to advanced stages.…”

The origins of bladder cancer

“…71 Likewise, microsatellite analysis of exfoliated urine to detect DNA LOH, combined with performance of UroVysion FISH and conventional urine cytology, enabled detection of over 93% of patients with recurrent bladder cancer. 72 Analysis of urine for microsatellite alterations shows a high correlation with invasive tumor resident within the bladder 73 and exhibits high sensitivity for patients who have invasive cancer. 74 Surprisingly, the most important biomarker of bladder cancer is microscopic or gross hematuria, which occurs in 85% of patients with bladder cancer.…”

Detection of Urothelial Bladder Cancer Cells in Voided Urine Can Be Improved by a Combination of Cytology and Standardized Microsatellite Analysis

“…Thirty-eight Urine Samples. As described previously (17), voided urine samples of 119 patients scheduled for transurethral resection were prospectively collected over a period of 20 months and matched with their corresponding biopsies. Of these, 81 biopsies proved to be neoplastic on histologic examination.…”

“…Overlapping cells were not analyzed. Samples were scored as FISH positive, if ≥4 cells showed at least 3 copies of any of the centromeric signals for chromosomes 3, 7, and 17 and if ≥12 cells displayed a homozygous loss of 9p21 (17). Due to technical reasons, only 2 of the 38 (5%) nonneoplastic urine samples were analyzable with FISH.…”

“…Microsatellite analysis of loss of heterozygosity (LOH) and FISH were among the most promising markers for surveillance (16). We and others have previously shown that the detection of bladder carcinoma cells can be improved by standardized microsatellite analysis (17,18). Over 93% of patients with recurrent bladder cancer disease were identified by a combination of microsatellite (LOH) analyses and cytology of their voided urine samples.…”