382 Prognostic impact of disseminated CK-20 positive tumour cells in bone marrow from bladder cancer patients after radical cystectomy

381 RT-PCR OF URINE SURVIVIN DIAGNOSIS OF TRANSITIONAL URINARY BLADDER uG.‘,

Passebosc-Fame

AND

K.2, Gentil-Perret

MN/CA9 FOR CELL CARCINOMA

A.3, Lambert

NON-INVASIVE (TCC)

OF

C.a, Genin C.2, Tostain J.’

382 PROGNOSTIC IMPACT OF DISSEMINATED TUMOUR CELLS IN BONE MARROW FROM PATIENTS AFTER RADICAL CYSTECTOMY Retz M.‘,

Lehmann

J.*, Rotering

J?, Eppelmann

CK-20 BLADDER

U.3, Stockle

POSITIVE CANCER

M.a

iCHU Saint Etienne, Department of Urology - Andrology, Saint Etienne, France, ZCHU Saint Etienne, Department of Clinical Immunology, Saint Etienne, France, QZHU Saint Etienne, Department of Pathology, Saint Etienne, France

‘UCSF, Department of Anatomy, San Francisco, United States, *Saarland University, Department of Urology and Paediatric Urology, HomburgiSaar, Germany, Ylniversity Kiel, Department of Urology, Kiel, Germany

INTRODUCTION & OBJECTIVES: Currently, the diagnosis of TCC of urinary bladder depends on cystoscopy or cytology. They are either invasive or lack of sensitivity The biomarker technology of gene diagnosis may improve the detection and monitoring of patients with TCC. Sun&in and MN/CA9 are two new markers that are expressed in tumour cells but not in normal cells. These two markers are also expressed in TCC of urinary bladder. The present study used the two markers for RT-PCR of urine for a non-invasive diagnosis of TCC of urinary bladder.

INTRODUCTION & OBJECTIVES: Systemic progression is the leading cause of death in bladder cancer patients after radical cystectomy. The aim of this present study was to assess the prognostic significance of circulating tumor cells detected by Cytokeratin 20 reverse-transcriptase polymerase chain reaction (CK-20 RTPCR) in bone marrow from bladder cancer patients.

MATERIAL & METHODS: 19 urine samples (> 50 ml) were collected sterilely before transurethral resection or cystectomy for bladder turnour. The diagnosis of TCC was established by the postoperative pathology. 8 urine samples from 6 non-TCC subjects and 2 patients with TCC after several days’ operation were used as control. The urine was centrifuged and washed once with hanks solution. The collected urine cells were lysed by RLT solution. The total RNA was extracted by a column. We designed specific primers for survivin and MNICA9. One-step RT-PCR was performed to check the presence of the gene markers. A cancer cell line was used as a positive control of RT-PCR. p-a&in was used to check the quality of the extraction of RNA. RESULTS: The distribution of tumour grade was: 3 grade I; 6 grade II and 10 grade III. The stage was: 8 pTa; 5 pT1; 3 pT2; 1 pT3a; 1 pT3b and 1 pT4. The extraction of total RNA from urine samples was successful in all cases. RT-PCR was positive in 15119 (79%) for survivin and in 10119 (53%) for MNICA9. When the two markers were combined, the positive RT-PCR of gene markers was 17/19 (90%). In contrast, there was no positive for either survivin or MN/CA9 in the control urine samples. CONCLUSIONS: This preliminary study demonstrates that RT-PCR of urine is feasible, and may provide a non-invasive, simple and sensitive diagnosis of TCC with a high specificity. Survivin and MN/CA9 could be used as the urine RT-PCR markers for the detection of tumour cells from TCC. A large scale study is needed to verify the sensitivity and specificity of this molecular assay and to evaluate its role for the diagnosis and monitoring of TCC patients.

383 HIGH RISK UPPER TRACT A MULTIVARIATE ANALYSIS Skolarikos

A.‘,

Griffths

TRANSITIONAL OF RISK FACTORS

L.a, Thomas

CELL

D.3, Neal D.4, Kelly

MATERIAL & METHODS: Bone marrow samples from 45 bladder cancer patients were drawn prior to radical cystectomy. The range of the tumour stages were pTl/CispNOMO n=7, pT2pNOMO n=14, pT3ipT4apNOMO n=13 and pT2pT4apNlipN2MO n=ll. CK-20 positive cells were detected by RT-PCR and analysed with respect to the endpoints of tumour progression and cancer death. CK-20 gene expression was also compared to clinical and histopathological parameters for prognostic significance by univariate and multivariate analysis (Cox regression). Clinical follow-up data were available on 45 patients (median: 19 months; range: l-58 months). RESULTS: 13 of 44 (29.5%) bladder tumour patients of all stages were positive for CK-20 detection in bone marrow. Progression free survival rate after 48 months was significantly shorter in the CK-20 positive group with 25.6% as compared to CK-20 negative patients with 58.3% (log-rank: p=O.O28). Furthermore, CK-20 positive tested patients had a significantly shorter tumour specific survival rate of 38.4% than CK-20 negative patient group with 64.6% (p=O.Ol). Multivariate analysis revealed that CK-20 and lymph node status provided independent prognostic information with respect to progression free survival. CONCLUSIONS: The detection of disseminated CK-20 positive tumour cells in bone marrow from bladder cancer patients is an independent prognostic factor and should be included in decisions concerning adjuvant therapies in early tumour stages. P22

PROSTATE CANCER: EPlDEMlOCOGY AND SCREENING Friday, 26 March, 12.15-13.45, Hall W Blue levet

CARCINOMA:

J.4

‘Athens Medical School, 2nd Department of Urology, Athens, Greece, *University of Lechester, Department of Urology, Lechester, United Kingdom, 3University of Newcastle Upon Tyne, Department of Urology, Newcastle Upon Tyne, United Kingdom, YJniversity of Cambridge Addenbrokes Hospital, Department of Urology, Cambridge, United Kingdom INTRODUCTION & OBJECTIVES: Conservation of the upper urinary tract is feasible in patients with low risk TCC (Ta GI or GII). The aim of this study was to determine the effectiveness of radiological, pathological and cytological investigative procedures to distinguish between high and low risk upper tract tumours. MATERIAL & METHODS: Between 1989 and 2001, 140 patients underwent nephroureterectomy for upper tract TCC. Pre-operative IVU (n=l17), ultrasound (n=89), retrograde pyelography (RP) (n=89), CT (n=Sl), ureteroscopic biopsy (n=51) and cytology (n=128) were evaluated without knowledge of the final pathological status. High risk TCC was defined as GIII or Tl-T4 in the pathological specimen. RESULTS: High risk TCC was associated with non-fimctionhydronephrosis on IVU (P=O.OOS); stricture on RP (P=O.O12); disease extending beyond the ureteric wallilymphadenopathy on CT (P=O.OOS); GII/GIII on biopsy (P=O.O02); and suspicious/positive cytology (P=O.O2). Hydronephrosismass on ultrasound and biopsy stage Tl-T4 were not associated with high risk TCC (Fisher’s exact test). In a multivariate logistic regression analysis, IVU (P=O.O04) and CT (P=O.Ol) were independent predictors of high risk TCC. RP did not add to IVU. In an analysis of biopsy grade and cytological status, only cytology was a significant predictor of high risk disease (P=O.O09). High risk status on IVU, RP, CT, biopsy grade, cytology detected 57%, 16 %, 44%, 94% and 54% respectively of the high risk ‘rumours. A combination of IVU, CT and cytology detected 79% (65182) of the high risk cases. The median follow-up was 26 months. Patients with high risk tumours died significantly earlier from TCC than those with low risk tumours (Cox regression; P=O.Ol). CONCLUSIONS: CT and IVU provide independent information in the prediction of high risk upper tract TCC. Cytological evaluation of exfoliated cells is a useful adjunct. European

Urology

Supplements

3 (2004)

No. 2, pp. 98

WITHDRAWN

384