Expression of p53 family genes in urinary bladder cancer: correlation with disease aggressiveness and recurrence.

p53 is a tumour suppressor gene with an established role in the majority of human neoplasias. Its homologues-p63 and p73-cannot be classified as tumour suppressors, since they encode isoforms with oncogenic properties as well. p63 plays a crucial role in epithelial cell differentiation and p73 is essential for neuronal cell development. The p63 and p73 expressions have been investigated in a variety of human tumours including bladder carcinomas; yet, this is the first study to simultaneously analyse the transcriptional levels of all p53 family members in bladder cancer. Using quantitative real-time polymerase chain reaction, we measured the mRNA expression of p53, p63 and p73 in 30 bladder tumours, each paired with adjacent normal tissue. All three studied genes were up-regulated in malignant specimens, p53 by 1.9-fold, p63 by threefold and p73 by twofold, respectively. Further analysis suggested that p63 and p73 act independently of p53 in the malignant bladder epithelium. Statistical analysis revealed that p63 overexpression was more frequent in recurrent bladder tumours (p = 0.045) and in older patients (p = 0.022). Papillary tumours also exhibited abnormal p63 expression (p = 0.026). Finally, p73 was up-regulated in Grade III one-site tumours (p = 0.040). Our results indicate that all p53 family members are abnormally expressed in bladder cancer but do not act synergistically. High levels of p63 correlate with non-muscle invasive tumours with frequent relapses, whereas p73 overexpression is associated with a more aggressive tumour phenotype.

Plasmacytoid urothelial carcinoma of the bladder: a rare malignancy.

In 2004, the WHO recognized the plasmacytoid subtype as a distinct variant of vesical malignancy. We present a case of plasmacytoid urothelial carcinoma of the bladder treated with radical surgery and adjuvant chemotherapy, thus achieving long-term survival. A 70-year-old woman presented with persistent dysuria and underwent cystoscopy which revealed the presence of diffuse deformity, involving the right lateral vesical wall. Histology revealed the presence of muscle-invasive urothelial carcinoma of the plasmacytoid variant. The patient subsequently underwent radical cystectomy and orthotopic ileal neobladder substitution, as well as adjuvant chemotherapy. At 36 months of follow-up, the patient is free of local recurrence and metastases, while her voiding function is well preserved. Bladder plasmacytoid urothelial carcinoma is considered a rare tumor, with unique microscopic and immunohistochemical features. The ideal therapeutic approach is debatable, but the combination of radical surgery and chemotherapy should constitute the mainstay of management.

Safety of ultrasound-guided transrectal extended prostate biopsy in patients receiving low-dose aspirin.

PURPOSE: To determine whether the peri-procedural administration of low-dose aspirin increases the risk of bleeding complications for patients undergoing extended prostate biopsies. MATERIALS AND METHODS: From February 2007 to September 2008, 530 men undergoing extended needle biopsies were divided in two groups; those receiving aspirin and those not receiving aspirin. The morbidity of the procedure, with emphasis on hemorrhagic complications, was assessed prospectively using two standardized questionnaires. RESULTS: There were no significant differences between the two groups regarding the mean number of biopsy cores (12.9 +/- 1.6 vs. 13.1 +/- 1.2 cores, p = 0.09). No major biopsy-related complications were noted. Statistical analysis did not demonstrate significant differences in the rate of hematuria (64.5% vs. 60.6%, p = 0.46), rectal bleeding (33.6% vs. 25.9%, p = 0.09) or hemospermia (90.1% vs. 86.9%, p = 0.45). The mean duration of hematuria and rectal bleeding was significantly greater in the aspirin group compared to the control group (4.45 +/- 2.7 vs. 2.4 +/- 2.6, p = < 0.001 and 3.3 +/- 1.3 vs. 1.9 +/- 0.7, p < 0.001). Multivariate logistic regression analysis revealed that only younger patients (mean age 60.1 +/- 5.8 years) with a lower body mass index (< 25 kg/m2) receiving aspirin were at a higher risk (odds ratio = 3.46, p = 0.047) for developing hematuria and rectal bleeding after the procedure. CONCLUSIONS: The continuing use of low-dose aspirin in patients undergoing extended prostatic biopsy is a relatively safe option since it does not increase the morbidity of the procedure.

Mutational analysis of the BRAF gene in transitional cell carcinoma of the bladder.

PURPOSE: Mutational activation of the MAP kinase pathway is frequently found in many types of cancer. Recently, activating mutations in the BRAF gene, an important activator of this pathway, have been described in several tumor types including melanoma, colorectal and papillary thyroid cancer. The most frequent mutation in exon 15 (V600E) as well as several other mutations within exons 11 and 15 result in constitutive activation of the oncoprotein. MATERIALS AND METHODS: Our study aimed to investigate BRAF mutations in 30 human bladder tumors and their adjacent normal tissues. The V600E mutation was screened by PCR/RFLP and exons 11, 14 and 15 of BRAF including intron-exon boundaries were sequenced. RESULTS: We detected two tumor specimens bearing two different mutations, both of which were found in exon 15. One sample showed the T1799A (V600E) and the other the G1798T (V600L) mutation. The first specimen was stage pT1a and grade II, whereas the second was stage pT2b and grade III. No mutations within the coding region of exons 11, 14, 15 and the intron-exon junctions for the remaining samples were found. CONCLUSIONS: Our results suggest that involvement of BRAF mutations in the development of transitional cell carcinoma of the bladder is infrequent.

Expression analysis of peptide growth factors VEGF, FGF2, TGFB1, EGF and IGF1 in prostate cancer and benign prostatic hyperplasia.

Peptide growth factors play an important role in several intracellular processes, such as cellular growth and differentiation, angiogenesis and apoptosis, as well as in carcinogenesis, since they contribute significantly to the malignant transformation. The prostate gland is abundant in growth factors. The two most known prostatic diseases, prostate cancer (PCa) and benign prostatic hyperplasia (BPH), are among the most common diseases that affect elderly men. This study was conducted using a quantitative real-time RT-PCR method in order to determine mRNA expression levels of peptide growth factors VEGF, FGF2, TGFB1, EGF, and IGF1 in tissue specimens from 42 patients with PCa, 42 with BPH, and 10 normal prostate samples obtained post-mortem from young individuals, in order to examine their association with prostatic hyperplasia and neoplasia. Our results show that in PCa, growth factors VEGF, EGF and FGF2 are overexpressed, while TGFB1 and IGF1 have reduced mRNA levels. In BPH, transcript levels of FGF2 and EGF are normal, while VEGF, TGFB1 and IGF1 exhibit downregulation. Further statistical analysis revealed that PCa patients with high levels of PSA blood levels have decreased FGF2 expression (p=0.016). Additionally, cancer patients with low Gleason score (<7) have increased EGF (p=0.035) and IGF1 (p=0.031) mRNA levels. IGF1 levels are also elevated in tumors with TNM stages T1-T2 (p=0.030). In BPH, older patients have reduced EGF expression (p=0.018), while IGF1 is overexpressed in younger patients (p=0.041). Additionally, the co-expression pattern of the five studied growth factors differs significantly among normal, benign and malignant prostate. These results implicate VEGF, FGF2, TGFB1, EGF and IGF1 in the development of both PCa and BPH, rendering them potential targets for disease detection, monitoring and therapy.

Deregulation of p73 isoform equilibrium in benign prostate hyperplasia and prostate cancer.

p73, a p53 homologue important for growth suppression, differentiation and induction of apoptosis, utilizes different promoters and undergoes alternative splicing to produce several isoforms differing in their ability to overlap p53 functions. Using reverse transcriptase-polymerase chain reaction (RT-PCR) to assess the mRNA levels of p53, p73 (total and isoforms specific for exons 2 and 13), MDM2, CDKN1A and beta2-microglobulin as internal control, we analyzed 35 prostate carcinomas and 44 benign prostate hyperplasias (BPH) compared to 14 normal prostates. Shift of p73 isoform mRNA levels from exon 13 lacking to exon 13 containing copies was observed in 80% of prostate cancer cases and in 52.3% of BPH specimens, and from exon 2 containing to exon 2 lacking (p73Deltaexon2) transcripts in 45.7% of cancer cases, but only in 9.1% of BPH samples. From these findings we deduce that p73 isoform balance is disrupted in prostate cancer and BPH, suggesting that this disequilibrium could play an important role in both prostate hyperplasia and malignancy.

Evaluation of neutrophil gelatinase-associated lipocalin, interleukin-18, and cystatin C as molecular markers before and after unilateral shock wave lithotripsy.

OBJECTIVE: To investigate the impact of shock wave lithotripsy (SWL) on renal tissues using neutrophil gelatinase-associated lipocalin (NGAL), cystatin C, and interleukin 18 (IL-18) levels in serum and urine and to examine the relationship of these biomarkers with patient and calculus characteristics as well as SWL treatment parameters. MATERIALS AND METHODS: Thirty-seven patients with renal calculi were included in this study. Blood and urine samples were attained from each patient at 4 time points; immediately before SWL, 6 hours after, 3 days after, and 10 days after the SWL. A new generation lithotripter was used for all cases. Serum and urine NGAL concentrations were measured using commercially available enzyme-linked immunosorbent assay kits according to manufacture's protocol. The concentration of cystatin C was measured in serum, whereas IL-18 concentration was assessed in urine. RESULTS: There were no statistically significantly differences in the levels of NGAL in serum and urine before and after SWL. The mean levels of cystatin C in serum appeared significantly higher 3 and 10 days after SWL. No statistically significant differences were identified between levels of IL-18 before and after SWL. Patients with diabetes mellitus demonstrated significantly higher baseline cystatin C levels. There was no correlation between calculus characteristics or treatment parameters and the levels of all 3 biomarkers after SWL. CONCLUSION: The results of this study indicate that SWL is associated with minimal acute injury to renal tissues. Our findings support the safety profile of new generation lithotripters, provided orthodox indications and treatment principles are followed.

Expression profile of CYP1A1 and CYP1B1 enzymes in colon and bladder tumors.

BACKGROUND: The cytochrome P450 CYP1A1 and CYP1B1 enzymes are involved in carcinogenesis via activation of pro-carcinogenic compounds to carcinogenic metabolites. CYP1A1 and CYP1B1 have shown elevated levels in human tumors as determined by qRT-PCR and immunohistochemical studies. However studies that have examined CYP1 expression by enzyme activity assays are limited. RESULTS: In the current study the expression of CYP1A1 and CYP1B1 was investigated in a panel of human tumors of bladder and colorectal origin by qRT-PCR and enzyme activity assays. The results demonstrated that 35% (7/20) of bladder tumors and 35% (7/20) of colon tumors overexpressed active CYP1 enzymes. CYP1B1 mRNA was overexpressed in 65% and 60% of bladder and colon tumors respectively, whereas CYP1A1 was overexpressed in 65% and 80% of bladder and colon tumors. Mean mRNA levels of CYP1B1 and CYP1A1 along with mean CYP1 activity were higher in bladder and colon tumors compared to normal tissues (p<0.05). Statistical analysis revealed CYP1 expression levels to be independent of TNM status. Moreover, incubation of tumor microsomal protein in 4 bladder and 3 colon samples with a CYP1B1 specific antibody revealed a large reduction (72.5 ± 5.5 % for bladder and 71.8 ± 7.2% for colon) in catalytic activity, indicating that the activity was mainly attributed to CYP1B1 expression. CONCLUSIONS: The study reveals active CYP1 overexpression in human tumors and uncovers the potential use of CYP1 enzymes and mainly CYP1B1 as targets for cancer therapy.

Prospective comparative study of endoscopic management of bladder lithiasis: is prostate surgery a necessary adjunct?

OBJECTIVES: To present the results of a prospective study comparing transurethral cystolithotripsy and simultaneous transurethral resection of the prostate (TURP), with transurethral cystolithotripsy and medical treatment of benign prostatic hyperplasia. The traditional dogma that bladder lithiasis constitutes an absolute indication for prostatic surgery has recently been questioned. METHODS: A total of 64 patients with bladder calculi were included in the present study. In all patients, stone clearance was achieved transurethrally. The patients in group 1 (n=32) underwent TURP during the same session, and the patients in group 2 (n=32) underwent medical therapy for benign prostatic hyperplasia (tamsulosin plus finasteride). RESULTS: The mean follow-up was 28.23±8.84 months. No statistically significant differences were found between the 2 groups regarding the preoperative parameters (age, International Prostate Symptom Score, prostatic volume, peak urinary flow rate, postvoid residual urine volume, prostate-specific antigen level, and bladder stone characteristics). Both groups experienced statistically significant postoperative improvements in the International Prostate Symptom Score, peak urinary flow rate, and postvoid residual urine volume. However, patients in group 1 experienced a more pronounced improvement in the International Prostate Symptom Score (P=.02) and peak urinary flow rate (P=.001). In total, 11 patients in group 2 underwent TURP during follow-up, with medical management considered to have failed. Multivariate logistic regression analysis revealed the postvoid residual urine volume as an independent risk factor that predicted the need for TURP in group 2 patients (odds ratio 1.033, 95% CI for odds ratio 1.007-1.060, P=.014). CONCLUSIONS: The findings of the present study have provided useful information on the natural history of bladder lithiasis, particularly in the context of improved patient consultation.

Safety of ultrasound-guided transrectal extended prostate biopsy in patients receiving low-dose aspirin

PURPOSE: To determine whether the peri-procedural administration of low-dose aspirin increases the risk of bleeding complications for patients undergoing extended prostate biopsies. MATERIALS AND METHODS: From February 2007 to September 2008, 530 men undergoing extended needle biopsies were divided in two groups; those receiving aspirin and those not receiving aspirin. The morbidity of the procedure, with emphasis on hemorrhagic complications, was assessed prospectively using two standardized questionnaires. RESULTS: There were no significant differences between the two groups regarding the mean number of biopsy cores (12.9 ± 1.6 vs. 13.1 ± 1.2 cores, p = 0.09). No major biopsy-related complications were noted. Statistical analysis did not demonstrate significant differences in the rate of hematuria (64.5 percent vs. 60.6 percent, p = 0.46), rectal bleeding (33.6 percent vs. 25.9 percent, p = 0.09) or hemospermia (90.1 percent vs. 86.9 percent, p = 0.45). The mean duration of hematuria and rectal bleeding was significantly greater in the aspirin group compared to the control group (4.45 ± 2.7 vs. 2.4 ± 2.6, p = < 0.001 and 3.3 ± 1.3 vs. 1.9 ± 0.7, p < 0.001). Multivariate logistic regression analysis revealed that only younger patients (mean age 60.1 ± 5.8 years) with a lower body mass index (< 25 kg/m2) receiving aspirin were at a higher risk (odds ratio = 3.46, p = 0.047) for developing hematuria and rectal bleeding after the procedure. CONCLUSIONS: The continuing use of low-dose aspirin in patients undergoing extended prostatic biopsy is a relatively safe option since it does not increase the morbidity of the procedure.