816 Chronic prostatitis — a nationwide survey of all urologists in Switzerland

816 CHRONIC PROSTATITIS GISTS IN SWITZERLAND Zbrun

S., Schumacher

University

Hospital

- A NATIONWIDE

SURVEY

M., Studer U.E., Hochreiter of Bern, Department

OF ALL

UROLO-

DUAL ROLE OF HUMAN URINARY

W.

of Urology,

P45 UROTHELIAL TUMOURS: BASIC RESEARCH II Saturday, 27 March, 15.15-16.45, Hall I/ Blue level CEACAMl BLADDER

Fernando M.‘, Oliveira-Ferrer Sevinc S.2, Kilic E.3, Friedrich

817

IN ANGIOGENESIS CANCER

AND

L.‘, Tilki D.2, Ziegeler G.2, Hauschild M.4, Wagener C.5, Ergiin S.4

INVASION

OF

.I.‘, Irmak

Bern, Switzerland

INTRODUCTION & OBJECTIVES: To document the current management of Chronic Prostatitis (CP) by urologists in Switzerland.

perception

and

MATERIAL & METHODS: In May 2003, we sent a questionnaire to all 154 urologists practicing in Switzerland at that time. The questionnaire consisted of 19 questions concerning prevalence, aetiology, diagnostic assessment. treatment and prognosis of CP. The returned questionnaires were statistically analyzed. RESULTS: Of the 154 questionnaires sent, 76 were returned (49%). Respondents see a median of 10 CP patients per month, 3 of them being newly diagnosed cases. This would correspond to a prevalence of255 and an incidence of 76 per 100’000 inhabitants for CP in Switzerland. Twenty-one percent of respondents believe in an infectious aetiology of CP. whereas 28% think that infection is not the aetiology of CP. For routine diagnostic assessment, the most commonly used tests are digital rectal examination, dipstick urine analysis, ultrasound for post-void residual urine assessment and microscopic urine analysis (91%, 75%, 73% and 67% of respondents, respectively). Microscopic and microbiologic analysis of post prostate massage urine (VB3) as well as symptom assessment by the National Institute of Health-Chronic Prostatitis Symptom Index NIH-CPSI are less frequently used (46%, 34% and 12% of respondents; respectively). The predominant treatment prescribed for CP is antibiotics (75%). Ninety-three percent of respondents who regard infection as aetiology of CP routinely use antibiotics as first line therapy. Surprisingly. also 80% of respondents who do not believe in an infectious aetiology prescribe antibiotics. When asked about the therapeutic consequences of VB-analysis, almost half of respondents would prescribe antibiotic treatment in the absence of leukocyturia. Tfeatments with non-steroidal antiinflammatory drugs or alpha-blockers are less frequently adopted as first line therapy (30% and 17% of respondents, respectively). The mean overall success rates of initial therapy reported (all treatments included) is 60%, but the mean recurrence rate after 6 months reported is 48%. CONCLUSIONS: One of 5 urologists believes in an infectious aetiology of CP. However, 3 of 4 urologists prescribe antibiotic treatment as first line therapy. Only a minority of respondents routinely perform prostate massage with analysis of VB3 when suspecting CP, and antibiotic treatment is often prescribed even without evidence of inflammation in VB3. To our knowledge, this is the first European study of this kind.

‘University Hospital Hamburg, Anatomy/ Urology, Hamburg, Germany, *University Hospital Hamburg, Anatomy, Hamburg, Germany. 3University Hospital Hamburg, Pathology, Hamburg, Germany. 4University Hospital Hamburg, Urology, Hamburg, Germany, 5University Hospital Hamburg, Clinical Chemistry, Hamburg, Germany INTRODUCTION & OBJECTIVES: Angiogenesis is a prerequisite for tumour growth and metastasis. It is regulated by a net balance of angiogenic activators such as the member of vascular endothelial growth factor (VEGF) family (VEGF-A, VEGF-C, VEGF-D) and angiogenic inhibitors including angiostatin and endostatin. Also. cell adhesion molecules play an essential role in the morphogenesis of newly sprouting blood vessels during turnour vascularization. Recently, we demonstrated that the carcinoembryonic antigen-related cell adhesion molecule-l (CEACAMI) acts pro angiogenic and as a morphogenetic effector for VEGF (Ergun et al., Mol. Cell, 2000). MATERIAL, METHODS &RESULTS: Here, we show that CEACAMl is expressed in umbrella cells of normal bladder urothelium, but it is down-regulated in superficial bladder cancer such as pTa and Tis. In contrast, CEACAMI is up-regulated in the endothelia of adjacent angiogenic blood vessels. To mimic the CEACAMl downregulation in normal bladder urothelium. CEACAM 1 was silenced in bladder cancer cell lines 486~ and RT4 using siRNA technique. CEACAMl down-regulation was confirmed at the protein level by Western blot analyses using the CEACAMI-specific antibody 4Dl/C2. In both bladder cancer cell lines, CEACAMI-silencing leads to a significant up-regulation of VEGF-D in real-time quantitative RT-PCR. Correspondingly, supematant from the CEACAMl-overexpressing bladder cancer cell line 486~ reduces, but that from CEACAMl-silencing increases VEGF-induced endothelial migration and tube formation. CEACAMl is present in angiogenic blood vessels of all invasive bladder cancer forms and in urine samples of patients with superficial (70%) and invasive (loo”/,) tumours. CONCLUSIONS: These data suggest a unique inverse angiogenic activity of CEACAMl depending on which cells expressing it. The enithelial down-regulation and endothelial uo-regulation of CEACAMl are involved in the switch from non-invasive and non-vascularized to invasive and vascularized bladder cancer. CEACAMI might be a useful diagnostic marker and a promising endothelial target for bladder cancer therapy.

818 GROWTH CANCER Nogawa

INHIBITION AND APOPTOSIS CELLS BY PLK-1 SIRNA M., Yuasa T., Kimura

S., Maekawa,

INDUCTION

OF BLADDER

819 CAN INHIBITION OF EPIDERMAL GROWTH FACTOR (EGFR) WITH GEFITINIB (‘IRESSA’, ZD1839) IMPROVE TO IONISING RADIATION IN BLADDER CANCER CELL

University

Hospital,

Transfusion

RECEPTOR RESPONSE LINES?

T. Colauhoun

Kyoto

S.‘,

Medicine

and Cell Therapy;

Kyoto,

A.J., Wozniak

S.P., Tulchinsky

E.M.,

Kriajevska

M., Mellon

J.K

Japan

INTRODUCTION & OBJECTIVES: Despite advances in medical oncology, radiation therapy, and molecular and cell biology, the mainstay in the management of invasive bladder cancer continues to radical cystectomy. From the point of improvement of quality of life, however, a new management to preserve the bladder has been desired. RNA interference is a newly discovered cellular pathway for the silencing of sequence-specific genes at the mRNA level by the introduction of the cognate double-stranded small interfering RNA (siRNA) and recently introduced into cancer therapy. Here we show growth inhibition and apoptosis induction of bladder cancer cells by polo like kinase-1 (PLK-1) siRNA. Expression of PLK-1, which has several functions in mitotic progression, is elevated in a broad range of human tumours. The aim of this study is to investigate the possibility as a therapeutic tool of PLK-I siRNA against bladder cancer. RilATERIAL & METHODS: We introduced four PLK-1 specific siRNAs into 12 bladder cancer cell line using cationic liposome and accessed the expressionreduction effect by Western blotting. The inhibitory effects of PLK-1 function were determined by Western blotting using cyclin Bl antibody and immunocytochemistry using alpha-tubulin antibody. Growth inhibitory effect, apoptosis induction, and cell cycle distribution were determined by a modified MTT assay, annexin V staining, and FACS analysis, respectively. RESULTS: Abundant expression of PLK-1 were demonstrated in all but one (T24) bladder cancer, whereas no or only scarce expression were detected in normal cells. Among four PLK-1 siRNAs we designed, the siRNA 1412 possessed the inhibitoriest potential against cognate gene expression. The siRNA 1412 suppressed PLK-1 expression dose- and time-dependent manner. Degradation of cyclin Bl and bipolar spindle formation were prevented by siRNA 1412. Moreover, time and dose dependently growth inhibition, M-phase arrest, and apoptosis induction by siRNA 1412 were confirmed in bladder cancer cells. CONCLUSIONS: PLK-1 siRNA demonstrated in vitro anti-cancerous activity that acted predominantly during M-phase and consequently causes M-phase arrest of cancer cells. To elucidate the efficacy of PLK-1 siRNA therapy in viva, we have established several orthotopic bladder cancer models. The efficacy and safety of PLK-1 siRNA for bladder cancer in viva will be verified in the murine bladder cancer models.

University Leicester,

of Leicester, Department United Kingdom

of Cancer

Studies

and Molecular

Medicine,

INTRODUCTION & OBJECTIVES: Approximately 70% of muscle-invasive bladder cancers express the epiderrnal growth factor receptor (EGFR) and expression is associated with poor prognosis. Recent evidence suggests that irradiation can stimulate EGFR resulting in activation of intracellular cytoprotective signalling cascades. Thus inhibition of EGFR function e.g. using a small molecule tyrosine kinase inhibitor, may be a means of enhancing the radioresponsiveness of bladder turnours. MATERIAL & METHODS: 6 bladder cancer cell lines with variable EGFR expression levels were treated with the tyrosine kinase inhibitor, gefitinib (‘Iressa’, ZD1839), ionising radiation (IR) or combined gefitinib/IR. Growth inhibition rates were assessed using both MTT and clonogenic assays. Growth inhibition rates determined with IR alone and combined gefitinib/IR were compared to calculate the dose enhancement ratio (DER) achieved with the use of gefitinib in each cell line. Subsequently. EGFR expression levels in each cell line were determined using Western blotting and related to the previously calculated DER. RESULTS: Treatment with gefitinib and IR alone resulted in dose-dependent growth inhibition in all 6 cell lines (IC,, gefitinib 0.18-2.48 KM). In all 6 cell lines assessed using clonogenic assay the combination of gefitinib and IR resulted in significantly greater growth inhibition than treatment with either gefitinib or IR alone (Student’s t test, p=O.OOl-0.04; DER 1.19-2.00). The enhancement of growth inhibition achieved using combined gefitinib and IR was significant in 5 of 6 cell lines assessed by MTT assay (Student’s t test; p=O.OOl-0.03; DER I .382.08). There was no correlation between EGFR expression levels and DER (Pearson’s correlation; clonogenic assay, ~0.3 18, p=O.539; MTT assay, r=0.206, ~~0.695). CONCLUSIONS: Radio sensitisation can be achieved in bladder cancer cell lines by the addition of gefitinib to ionising radiation. Repetition of these in vitro findings using an in vI’~.o model (work currently underway) would provide sound scientific justification for translating this work into a clinical trial. ‘Iressa’ is a trademark of the AstraZeneca group of companies. European

Urology

Supplements

3 (2004)

No. 2, pp. 207