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MP-04.02: Enzymatic immuno-assay quantification of C-erbB-2 oncogene expression in urinary bladder cancer
MODERATED POSTER SESSIONS
MP-04: Bladder Cancer Monday, September 3 10:15-12:15 MP-04.01 Molecular mechanisms of the urinary bladder carcinogenesis (the intercellular proteolysis alteration) influenced by persistent, long-term, low-dose ionizing radiation in humans Romanenko AM1, Vozianov AF2, Fukushima S3 1 Department of Pathology; 2 Department of Urology, Institute of Urology, Academy of Medical Sciences of Ukraine, Kiev, Ukraine; 3Department of Pathology, Osaka City University Medical School, Osaka, Japan Objectives: Discovery of the intercellular ubiquitin-proteasomal proteolysis system was awarded by the Nobel Prize for chemistry in 2004. The incidence of urinary bladder cancer in the Ukraine, increased from 26.2 to 50.3 per 100, 000 population between 1986 and 2006, after the Chernobyl accident. Cesium 137 accounts for 90% of the internal radioactivity in the Ukrainian population and it is known to be eliminated via the urine. The present study was conducted to determine whether ubiquitination and sumoylation processes in cell proteolysis are altered in urinary bladder urothelium by chronic, long-term persistent low doses of ionizing radiation (IR) in male patients (with urinary retention) with benign prostate hyperplasia (BPH) and females with chronic cystitis living more than 20 years in Cesium 137 contaminated areas after the Chernobyl accident in Ukraine. Methods: Bladder urothelial biopsies from 45 patients who underwent suprapubic prostatectomy for BPH, as well as 20 specimens from 5 females with chronic cystitis, were subjected to histopathological and immunohistochemical studies of ubiquitin (Ub), the small Ub-related modifier 1 (SUMO1), SUMO E2 conjugating enzyme Ubc9, and the cell cycle inhibitors p53 and p27Kip1. Results: Chronic proliferative atypical cystitis (“Chernobyl cystitis”), featuring multiple foci of dysplasia, and carcinoma in situ (CIS) were observed in 23 (92%) and 19 (76%), respectively, of 25 group 1 patients from radio-contaminated areas, in addition to one small pTa grade 1 urothelial carcinoma. Chronic cystitis with areas of mild dysplasia and urothelial hyperplasia were detected in 2 (10%) and 3 (15%)
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of 20 patients in the control group 2 from so-called clean (without radio-contamination) areas of Ukraine. Strongly elevated levels of Ub, SUMO1, Ubc9 and p53 as well as decreased levels of p27Kip1, were evident for patients in group 1, as compared to group 2 (all p ⬍ 0.001). Conclusion: Activation of ubiquitination and sumoylation processes in Chernobyl cystitis might be a compensatory reaction in response to the insufficient proteolysis of aberrant p53 and, on the other hand, lead to unscheduled degradation of p27Kip1 cell cycle negative regulator occurred due to the long-term low dose rate, IR exposure that may contribute to urothelial carcinogenesis caused by IR. MP-04.02 Enzymatic immuno-assay quantification of C-erbB-2 oncogene expression in urinary bladder cancer Mehena AA1, Wishahi M1, Hassan S2, ElSayed N3 1 Theodor Bilharz Research Institute, Urology Department, Heliopolis, Egypt; 2 Medical Chemistry Department, National Research Centre; 3 Zoology Oncology Unit, Faculty of Medicine, Ein Shams University, Cairo, Egypt Introduction: Over expression of several oncogenes, including c-erbB-2, have been implicated in bladder cancer biology. Methods: Using enzymatic immuno-assay (EIA) technique, a method that allows for a quantitative determination of c-erbB-2 expression level in tissues, we examined this oncogene for amplification in 69 bladder cancer tissue samples. The results were compared to those of bilharzial nonmalignant (5 cases) and control (20 cases) groups. Results: The c-erbB-2 mean value recorded in the control group was 0.8⫾0.08. It was expressed more often in the bilharzial non-malignant group (2.30⫾0.42) and in bladder cancer group either bilharzial associated (mean value 3.53⫾0.52) or non-bilharzial-associated (mean value 4.96⫾0.58). Statistical correlation revealed a high statistically significant difference (P⬍0.001) between the different groups. At a cut off value⬎1.9 (which was the highest normal value recorded), the positivity rate was higher in non bilharzial bladder cancer (82.1%) than bilharzial bladder cancer (60%) reach statistically significant level at P⬍0.05. In correlation between transitional and squamous cell types, the c-erbB-2 mean values were 4.45⫾0.50 in transitional and 4.14⫾0.70 in squamous cell types, with a statistically significant difference
(P⬍0.001). The positivity rate (77.3% in the transitional and 68% in the squamous cell carcinomas) lacks such significant difference (P⬍0.05). Statistical analysis also revealed statistically significant correlations between the lymph node status of the bladder cancer patients and the c-erbB-2 mean values (P⬍0.001), the positivity rate (P⬍0.05) being higher in the lymph node negative cases (4.79⫾0.54 and 84.2%) than the lymph node positive cases (3.81⫾0.59 and 61.3%).On the other hand, there was no statistically significant correlation between c-erbB-2 expression and pathological grades and tumor stages. Conclusion: These data indicated that c-erbB-2 oncogenes is expressed as an early event in bilharziasis and its over expression increases with tumorigenesis. It may be a diagnostic marker, but it has no prognostic value in assessing tumor progression and differentiation. MP-04.03 The value of tissue polypeptide antigen (TPA) in the after care patients with bladder cancer: relationship with conventional urine cytology Bantis A1, Zissimopoulos A2, Kalaitzis C1, Sountoulides P1, Giannakopoulos S1, Patris E1, Aggelonidou E2, Voudalikakis C1, Touloupidis S1 1 Urology Department, 2Nuclear Medicine Department, University Hospital, Alexandroupolis, Greece Introduction: The prognosis of bladder cancer has to be considered poor. Tissue polypeptide antigen (TPA) is present in the proteolytic fragments of cytokeratins 8, 18 and 19 as a component of the cytoskeleton of nonsquamous epithelia. The aim of this study were to determine the clinical usefulness of TPA in ser and urine as a tumour marker for the diagnosis of bladder cancer in comparison with conventional urine cytology. Methods: Freshly voided urine and blood samples obtained from 108 patients who presented for either urinary symptoms (30), for follow up of bladder tumour (45 invasive and 63 superficial) and 50 health individuals (blood donors) to estimate the cut of value were studied. In all cases cystoscopy or biopsies have been performed. Results: The cut – off value was 95U/ml (94⫾4,6). For superficial bladder cancer we found elevated TPA levels in 40% in serum and 35% in urine. Elevated TPA levels was also found in invasive bladder cancer 52% and 53% in serum and urine respectively. The sensitivity for all studied cases of TPA was 50% in serum and 39% in urine and the
UROLOGY 70 (Supplment 3A), September 2007
MP-04: Bladder Cancer Monday, September 3 10:15-12:15 MP-04.01 Molecular mechanisms of the urinary bladder carcinogenesis (the intercellular proteolysis alteration) influenced by persistent, long-term, low-dose ionizing radiation in humans Romanenko AM1, Vozianov AF2, Fukushima S3 1 Department of Pathology; 2 Department of Urology, Institute of Urology, Academy of Medical Sciences of Ukraine, Kiev, Ukraine; 3Department of Pathology, Osaka City University Medical School, Osaka, Japan Objectives: Discovery of the intercellular ubiquitin-proteasomal proteolysis system was awarded by the Nobel Prize for chemistry in 2004. The incidence of urinary bladder cancer in the Ukraine, increased from 26.2 to 50.3 per 100, 000 population between 1986 and 2006, after the Chernobyl accident. Cesium 137 accounts for 90% of the internal radioactivity in the Ukrainian population and it is known to be eliminated via the urine. The present study was conducted to determine whether ubiquitination and sumoylation processes in cell proteolysis are altered in urinary bladder urothelium by chronic, long-term persistent low doses of ionizing radiation (IR) in male patients (with urinary retention) with benign prostate hyperplasia (BPH) and females with chronic cystitis living more than 20 years in Cesium 137 contaminated areas after the Chernobyl accident in Ukraine. Methods: Bladder urothelial biopsies from 45 patients who underwent suprapubic prostatectomy for BPH, as well as 20 specimens from 5 females with chronic cystitis, were subjected to histopathological and immunohistochemical studies of ubiquitin (Ub), the small Ub-related modifier 1 (SUMO1), SUMO E2 conjugating enzyme Ubc9, and the cell cycle inhibitors p53 and p27Kip1. Results: Chronic proliferative atypical cystitis (“Chernobyl cystitis”), featuring multiple foci of dysplasia, and carcinoma in situ (CIS) were observed in 23 (92%) and 19 (76%), respectively, of 25 group 1 patients from radio-contaminated areas, in addition to one small pTa grade 1 urothelial carcinoma. Chronic cystitis with areas of mild dysplasia and urothelial hyperplasia were detected in 2 (10%) and 3 (15%)
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of 20 patients in the control group 2 from so-called clean (without radio-contamination) areas of Ukraine. Strongly elevated levels of Ub, SUMO1, Ubc9 and p53 as well as decreased levels of p27Kip1, were evident for patients in group 1, as compared to group 2 (all p ⬍ 0.001). Conclusion: Activation of ubiquitination and sumoylation processes in Chernobyl cystitis might be a compensatory reaction in response to the insufficient proteolysis of aberrant p53 and, on the other hand, lead to unscheduled degradation of p27Kip1 cell cycle negative regulator occurred due to the long-term low dose rate, IR exposure that may contribute to urothelial carcinogenesis caused by IR. MP-04.02 Enzymatic immuno-assay quantification of C-erbB-2 oncogene expression in urinary bladder cancer Mehena AA1, Wishahi M1, Hassan S2, ElSayed N3 1 Theodor Bilharz Research Institute, Urology Department, Heliopolis, Egypt; 2 Medical Chemistry Department, National Research Centre; 3 Zoology Oncology Unit, Faculty of Medicine, Ein Shams University, Cairo, Egypt Introduction: Over expression of several oncogenes, including c-erbB-2, have been implicated in bladder cancer biology. Methods: Using enzymatic immuno-assay (EIA) technique, a method that allows for a quantitative determination of c-erbB-2 expression level in tissues, we examined this oncogene for amplification in 69 bladder cancer tissue samples. The results were compared to those of bilharzial nonmalignant (5 cases) and control (20 cases) groups. Results: The c-erbB-2 mean value recorded in the control group was 0.8⫾0.08. It was expressed more often in the bilharzial non-malignant group (2.30⫾0.42) and in bladder cancer group either bilharzial associated (mean value 3.53⫾0.52) or non-bilharzial-associated (mean value 4.96⫾0.58). Statistical correlation revealed a high statistically significant difference (P⬍0.001) between the different groups. At a cut off value⬎1.9 (which was the highest normal value recorded), the positivity rate was higher in non bilharzial bladder cancer (82.1%) than bilharzial bladder cancer (60%) reach statistically significant level at P⬍0.05. In correlation between transitional and squamous cell types, the c-erbB-2 mean values were 4.45⫾0.50 in transitional and 4.14⫾0.70 in squamous cell types, with a statistically significant difference
(P⬍0.001). The positivity rate (77.3% in the transitional and 68% in the squamous cell carcinomas) lacks such significant difference (P⬍0.05). Statistical analysis also revealed statistically significant correlations between the lymph node status of the bladder cancer patients and the c-erbB-2 mean values (P⬍0.001), the positivity rate (P⬍0.05) being higher in the lymph node negative cases (4.79⫾0.54 and 84.2%) than the lymph node positive cases (3.81⫾0.59 and 61.3%).On the other hand, there was no statistically significant correlation between c-erbB-2 expression and pathological grades and tumor stages. Conclusion: These data indicated that c-erbB-2 oncogenes is expressed as an early event in bilharziasis and its over expression increases with tumorigenesis. It may be a diagnostic marker, but it has no prognostic value in assessing tumor progression and differentiation. MP-04.03 The value of tissue polypeptide antigen (TPA) in the after care patients with bladder cancer: relationship with conventional urine cytology Bantis A1, Zissimopoulos A2, Kalaitzis C1, Sountoulides P1, Giannakopoulos S1, Patris E1, Aggelonidou E2, Voudalikakis C1, Touloupidis S1 1 Urology Department, 2Nuclear Medicine Department, University Hospital, Alexandroupolis, Greece Introduction: The prognosis of bladder cancer has to be considered poor. Tissue polypeptide antigen (TPA) is present in the proteolytic fragments of cytokeratins 8, 18 and 19 as a component of the cytoskeleton of nonsquamous epithelia. The aim of this study were to determine the clinical usefulness of TPA in ser and urine as a tumour marker for the diagnosis of bladder cancer in comparison with conventional urine cytology. Methods: Freshly voided urine and blood samples obtained from 108 patients who presented for either urinary symptoms (30), for follow up of bladder tumour (45 invasive and 63 superficial) and 50 health individuals (blood donors) to estimate the cut of value were studied. In all cases cystoscopy or biopsies have been performed. Results: The cut – off value was 95U/ml (94⫾4,6). For superficial bladder cancer we found elevated TPA levels in 40% in serum and 35% in urine. Elevated TPA levels was also found in invasive bladder cancer 52% and 53% in serum and urine respectively. The sensitivity for all studied cases of TPA was 50% in serum and 39% in urine and the
UROLOGY 70 (Supplment 3A), September 2007