Prediction of recurrence in low and intermediate risk non-muscle invasive bladder cancer by real-time quantitative PCR analysis: cDNA microarray results.

The aim of the study was to define specific genetic profile in Ta and T1 urinary bladder carcinoma patients with and without recurrence by gene expression microarrays. Eleven patients with the time to recurrence shorter than one year (patients with recurrence) and 11 patients with time to recurrence longer than 4 years (patients without recurrence) were enrolled. Data from microarrays were subjected to a panel of statistical analyses to identify bladder cancer recurrence-associated gene signatures. Initial screening using the GeneSpring and Bioconductor software tools revealed a putative set 47 genes differing in gene expression in both groups. After the validation, 33 genes manifested significant differences between both groups. The significant expression was observed in the group of patients without recurrence by 30 genes of which the highest differences were detected by ANXA1, ARHGEF4, FLJ32252, GNE, NINJ1, PRICKLE1, PSAT1, RNASE1, SPTAN1, SYNGR1, TNFSF15, TSPAN1, and WDR34. These genes code for signal transduction, vascular remodeling and vascular endothelial growth inhibition mainly. In the group with recurrence, 3 genes had significant differences, the highest differences were identified by two genes (PLOD2 and WDR72). Loci of genes with significant changes of gene expression were located on characteristic chromosomes for bladder cancer: 7 loci on chromosome 9, 8 loci on chromosomes 1, 2, 3, 12,14,15,16, and 22. We have selected and validated 15 genes that are differentially expressed in superficial bladder cancer. We hope that this cohort of genes will serve as a promising pool of candidate biomarkers for early stage bladder cancer. Our results indicate that it may be possible to identify patients with a low and high risk of disease recurrence at an early stage using a molecular profile.

[Cystectomy in the treatment of bladder cancer]. / Cystektomie v léce nádoru mocového mechýre.

Radical cystectomy with pelvic lymphadenectomy and urinary diversion is an important component in the treatment of bladder cancer. It is considered the most important method in the therapy for muscle invasive and selected high-risk non-muscle invasive tumours with excellent local control and high complete remission rate. It consists of complete removal of tumour tissue in the bladder, small pelvis and regional lymph nodes. In males, urinary bladder and prostate are routinely removed; in females, bladder, uterus and anterior vaginal wall are removed. Urethrectomy is indicated only in selected situations. An integral part of the operation is the bilateral pelvic lymphadenectomy. Extravesical disease extension and lymph node positivity are unfavourable prognostic factors. Better prognosis is expected in patients with less than 5 positive nodes. Important prognostic factor is also the number of removed lymph nodes, which is a strong argument for meticulous bilateral pelvic lymphadenectomy. Lymph node density (number of positive nodes/ number of removed nodes) is considered as very important prognostic factor. Better prognosis can be expected in patients with less positive and more removed nodes.

[The p53 positivity in non-tumor mucosa in patients with superficial urinary bladder cancer]. / P53 pozitivita v nenádorove sliznici u pacientu s povrchovým uroteliiálním karcinomem mocoveho mechyre.

BACKGROUND: The aim of the study is to assess the prognostic value of p53 positivity in the non-tumor mucosa of urinary bladder in patients with superficial urinary bladder carcinoma. METHODS AND RESULTS: In 45 patients cold cup non-tumor mucosa samples were taken at the same time with the TUR of superficial urinary bladder carcinoma prospectively. Monoclonal antibody BP53-12-1 was used for the detection of p53 protein. When identifying positive colouring only the nuclear immunoreactivity was being evaluated. 200 nuclei at minimum were examined in several representative fields. The McCarthy method in Bacus modification was used to analyse the findings. It is a semiquantitative method which detects not only the percentage of p53 positive cells but also the intensity of positivity classified into four degrees (0 - negative, 1 - slightly positive, 2 - distinctly positive, 3 - strongly positive). The intensity of p53 positivity was quantified as HSCORE, where HSCORE = 7Pi (i + 1), in which i is one of the four see above degrees and Pi fluctuates from 0 % to 100 %. The result is a numerical figure from 100-400. A negative finding is of HSCORE 100, HSCORE of 400 is the highest possible. The samples were analysed in the analytical system LUCIA. The borderline value was quantified to HSCORE 200. All patients were carefully followed up and treated using usual schemes. The results were evaluated by the use of SAS system (Cary, USA). Thirty patients recurred during the follow-up and 7 of them progressed. The average HSCORE in those who did not recur was 130.2, in patients with the recurrence of tumor it was 162.5 and in patients with progression it was 169.2. We have found a correlation between the HSCORE and the risk of recurrence, which was statistically significant. CONCLUSIONS: The p53 positivity in non-tumor mucosa of urinary bladder in patients with superficial bladder cancer may bring additional information when predicting the risk of recurrence. More extensive studies need to be carried out.

Urinary cytology and quantitative BTA and UBC tests in surveillance of patients with pTapT1 bladder urothelial carcinoma.

OBJECTIVES: To compare results of urinary cytology, quantitative detection of human complement factor H-related protein (BTA TRAK), and urinary fragments of cytokeratins 8 and 18 (UBC IRMA) with the recurrence status in patients with pTapT1 bladder cancer and to define the possible role of these methods in a surveillance protocol. METHODS: We collected urine from 88 consecutive patients with primary pTapT1 tumors before the first transurethral resection (TURB) and before each follow-up cystoscopy. In all samples urinary cytology and quantitative BTA and UBC tests were performed. We compared results with recurrence status and with tumor characteristics in the case of recurrence. We constructed receiver operator characteristic (ROC) curves for quantitative methods. In addition, we evaluated individual cutoffs based on pretreatment levels. RESULTS: During the mean follow-up of 16.96 months, we performed 313 cystoscopies, 93 of which were positive in 51 patients. The sensitivity and specificity of cytology, BTA, and UBC were 19.8% and 99%, 53.8% and 83.9%, and 12.1% and 97.2%, respectively. The sensitivity of pTis detection was 66.6%, 0%, and 100%, respectively. With cutoffs set to a sensitivity of 90%, the specificity of BTA and UBC dropped to 24.8% and 20.4%, respectively. Individually calculated cutoffs did not provide a significant benefit. CONCLUSIONS: Because of high specificity and sensitivity in pTis detection, urinary cytology fulfills requirements for an adjunctive method to cystoscopy. Quantitative BTA and UBC tests have a low sensitivity in the detection of bladder cancer recurrence and cannot be used routinely to reduce the number of cystoscopies during follow-up.