Prostate cancer: Serum markers (II)

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adjuvant chemotherapy. For the entire cohort of patients, the mean follow-up was 2.6 years, and 47% are alive without disease, while 2% are alive with disease. In terms of cancer deaths, 30% are dead of disease and 4% are dead of other causes. The disease status of 17% is unknown. The overall mean survival was 2.76 years (CI 2.5-3.0, Kaplan-Meier). Despite a significantly shorter survival when compared to node negative patients, the mean survival for patients with N + was still 2.3 years (CI 1.86-2.84). In the N+ patients, there was no significant difference in survival between those who received an ileal conduit and those who received an orthotopic neobladder (p=0.265). CONCLUSIONS: Radical cystectomy remains the most effective form of local control among patients with invasive urothelial carcinoma and affords modest survival advantage even among those with N+ nodes. The type of urinary diversion does not affect these patients' survival. Source of Funding: None.

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these RCT's should be done, however, the magnitude of overall survival benefit with current combination chemotherapy regimens is likely to be small. We hypothesize that this reflects genetic heterogeneity in chemosensitivity of TCC; limiting the benefits of NAe. Efforts to identify and limit use of NAC to those who benefit are recommended. Source of Funding: None.

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EARLY METASTATIC PROGRESSION OF BLADDER CARCINOMA.MOLECULAR PROFILE OF PRIMARY TUMOR AND SENTINEL LYMPH NODE Amir Sherif>, Per-Uno Malmstrom,

FINAL RESULTS OF THE PROSPECTIVE EUROPEAN MULTICENTER STUDY OF COMPLEX PSA AND COMPLEX PSA INDICES FOR EARLY PROSTATE CANCER DETECTION

Zhi-Ping Ren, Manuel De La Torre, Kenneth Wester, Magnus Thorn, Uppsala, Sweden

Bob Djavan*, Alexandre Zlotta, Vincent Ravery, Peter Hammerer, Amir Kaisary, Piotr Dobronski, Andreas Reissigl, Mesut Remzi, Claude Schulman, Laurent Boccon Gibod, Michael Marberger, Vienna, Austria

INTRODUCTION AND OBJECTIVE: We characterized early metastatic progression of bladder carcinoma from the primary tumor, separated in the central part and invasive front, to the first lymphatic metastasis. METHODS: Included in this study were 8 patients undergoing sentinel lymph node detection for invasive bladder cancer, of whom 4 had metastasis in the sentinel lymph node and 4 were randomly chosen without metastases. After microdissection p53 genomic structure and immunohistochemical expression of p53,pRB,Ki67 and E-cadherin were analyzed. Microvessel density and apoptosis were also assessed. RESULTS: In 5 patients there were p53 gene mutations in the primary tumor, while 3 had the wild-type gene. The genotypes were identical in the central part and invasive front. All sentinel lymph node metastases harbored p53 mutations, in contrast to all nonmetastatic sentinel lymph nodes. Two patients had the same mutation as the primary tumor and one had an additional mutation. In a patient with a wild-type gene in each compartment of the primary tumor a mutation appeared in the corresponding sentinel lymph node metastasis. There was poor concordance of p53 mutation with protein status. The expression of p53,pRB,Ki67 ,E-cadherin, and the evaluation ofapoptosis and angiogenesis showed in most cases only slight variations in tumor compartments and the sentinel lymph node. CONCLUSIONS: In this study invasive bladder carcinoma involved monoclonal proliferations with a mainly homogenous biomarker profile. The first metastases in sentinel lymph nodes had a similar molecular profile but in half of the cases signs of clonal evolution appeared. Source of Funding: None.

1432 ROLE OF NEOADJUVANT CHEMOTHERAPY (NAC) IN TRANSITIONAL CELL CARCINOMA (TCC): A SYSTEMATIC REVIEW Joseph L Chin*, Eric W Winquist, London, ON, Canada; Roanne J Segal, Ottawa, ON, Canada; Himanshu R Lukka, Hamilton, ON, Canada INTRODUCTION AND OBJECTIVE: Although cisplatin-based combination chemotherapy has improved survival in metastatic TCC, the benefit of NAC in Stage II and III TCC prior to definitive therapy is unclear, based on all available reports. Moreover, the quality of evidence from these reports varies widely. A comprehensive and systematic review of the current evidence was undertaken to elucidate this issue. METHODS: A comprehensive search of electronic databases and handsearching of meeting abstracts, trial bibliographies and textbooks was undertaken. Only randomized controlled trials (RCT's), meta-analyses and systemic reviews were eligible, and only reports of ones providing comparisons of overall survival and/or disease-specific survival between NAC and local therapy alone were included. RESULTS: 13 RCT's and I individual patient data meta-analysis satisfied the inclusion criteria. The meta-analysis included only the 4 earliest RCT's and reported a mortality hazard ratio (HR) of 0.91 favoring NAC, which was not statistically significant (p=0.33). 10 of 12 RCT's reporting survival data showed no statistical significance of NAC on overall survival, including the largest study of NAC conducted to date (976 patients). In this trial,mortality was reduced by NAC (HR 0.85; 95% e.r., p=0.075) with an absolute difference in 3-year overall survival of 5.5%, but the effect did not reach statistical significance. As only 7 of the 13 RCT's provided published data suitable for statistical pooling, meta-analysis was not attempted and an interpretative summary was done. CONCLUSIONS: NAC should not be routinely offered prior to cystectomy and lor radiotherapy for stage II and III TCe. Individual patient meta-analysis of

INTRODUCTION AND OBJECTIVE: Complex PSA assays have been recently introduced as a new tools for early prostate cancer detection in patients with low (2.5 to 4) and intermediate (4 to 10 ng/mL) PSA levels. Two multicenter trials, the 7 site US trial and the European trial were designed to evaluate the performance of cPSA (US and European trial) and cPSA indices (European trial)including cPSAD, cPSATZ, cit PSA ratio, serum total PSA, PSAD, PSA-TZ and %free PSA. METHODS: This prospective multicenter European study included a total of 992 men with a serum total PSA between 4 ng/ml and 10 ng/ml. Transrectal ultrasound (TRUS)guided sextant biopsy and two additional transition zone biopsies were obtained in all patients. Biopsy outcome was correlated to the following parameters: serum total PSA, Complex PSA (Bayer Corporation, USA), cPSAD, cPSATZ, cit PSA ratio, c/f PSA ratio, fit PSA, PSAD and PSATZ. In addition to standard statistics a neural network (ANN) was used in the analysis with an advanced multiplayer perceptron selected for accuracy by a genetic algorithm. RESULTS: In those patients with 'IPSA 4 to 10 ng/mL, 33.8% were found to have prostate cancer and 66.1% were found to have a non-cancer histology. For men with intermediate PSA levels the respective Areas under the curve (AUCs) were: cPSA = 66.4%, cit PSA = 67.1 %, fit PSA = 66.4%, fPSA = 66.8 ng/mL, cPSAD = 62.2 %, PSAD = 42.9 %, PSATZ = 64.9 %, and cPSATZ = 67.8. CONCLUSIONS: In the intermediate PSA range (4 to 10 ng/mL), the performance of cPSA alone was identical to % free PSA, whereas cit PSA ratio and cPSATZ were slightly superior without reaching statistical significance. In this PSA range, cPSAD and clf PSA ratio were of no additional value even when an ANN was employed. Complex PSA and cPSA indices are certainly helpful in improving cancer detection in patients with PSA levels form 4 to 10 ng/mL without however outperforming %free PSA in terms of statistical performance. Their foremost advantage is however, easy handling and high molecular stability. Source of Funding: None.

1434 A NOVEL ASSAY FOR NICKED, MULTI-CHAIN FREE PSA ENHANCES DISCRIMINATION OF PROSTATE CANCER CASES FROM MEN WITH NO CANCER AND PSA-LEVELS OF 2-10 NGIML Pauliina M Niemela*, Turku, Finland; Thomas Steuber, Hartwig Huland, Hamburg, Germany; Hans Lilja, Malmo, Sweden; Kim Pettersson, Turku, Finland INTRODUCTION AND OBJECTIVE: Mildly elevated PSA-Ievels in blood (e.g. 2-10 ng/mL) increase the risk for prostate cancer but have low specificity for cancer (20-30%). Analysis of free vs. complex PSA (%fPSA) enhances discrimination of prostate cancer (PCa) from benign prostate disease. We recently reported that proportion of single-chain free PSA-subfractions are significantly higher in PCa compared to patients with no cancer. We now developed the first assay to measure nicked, multi-chain free PSA-forms containing an internal cleavage at Lys'4,-Lys'46 and evaluated the utility of this tool to discriminate cancer from non-cancer cases. METHODS: An antibody binding to Lys14,-Lys146 region in PSA is used to block free PSA-forms lacking internal cleavage at Lys14,-Lys146' Internally cleaved, nicked free PSA-forms (fPSA-N) are subsequently measured by an immunofluoroassay with a detection limit of 0.05 ng/ml. Anti-coagulated EDTAplasma was processed rapidly and frozen in one hour from venipuncture from men with PSA-Ievels of 2-10 ng/mL referred for sextant prostate biopsy. There were plasma from 52 men with biopsy-confirmed PCa and 55 men with either negative

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biopsy or no cancer at TURP. We also measured levels of total and free PSA and calculated complexed PSA from these parameters. RESULTS: Levels of fPSA-N were significantly higher in men with no cancer (0.4 ng/ml) compared to PCa-cases (0.24 ng/ml; p
1435 THE DIAGNOSTIC IMPACT OF PRO-PSA IN THE EARLY DETECTION OF PROSTATE CANCER (PCA) IN PATIENTS WITH A TOTAL PSA RANGE OF 4.0 TO 10.0 NGIML Robert W Veltri*, Masood A Khan, Baltimore, MD; Harry G Rittenhouse, Stephen D Mikolajczyk, Brea, CA; Lori J Sokoll, Daniel W Chan, Alan W Partin, Baltimore, MD INTRODUCTION AND OBJECTIVE: In contemporary screening populations, a major draw back of PSA is its relative lack of specificity especially in the diagnostic range of 4 to 10 ng/ml where this result may indicate either PCa approximately 30% of the time or benign disease about 70% of the time. ProPSA is a derivative of free-PSA (fPSA) consisting of the truncated forms (e.g. [-2]pPSA, [-4]pPSA) or the full length [-7]pPSA. There is growing evidence that proPSA is associated preferentially with prostate cancer. The objective of this study was to determine whether proPSA can positively influence the detection of early prostate cancer through reducing false positives. METHODS: Archival serum samples obtained from 93 men that underwent a systematic 12 core biopsy procedure at the Johns Hopkins Hospital (total PSA range 4.0 to 10.0 ng/mL) were assayed for %-free, total, and the three forms of proPSA (Hybritech Tandem Assays). Free PSA and cumulative sum of the individual proPSA forms ([-2], [-4] and [-7]) and their derivatives were determined. Prostate volumes were measured at the time of systematic 12-core biopsies for the detection of prostate cancer. The results were analyzed using univariate and multivariate logistic regression (LR) non parametric statistical methods. RESULTS: Of the 93 men assessed, 41 (44%) had evidence of prostate cancer (76% Gleason 5/6,19% Gleason 7, 5% Gleason 8). Using univariate LR total-PSA, fPSA, % fPSA, % sum-pPSA, and prostate volume all significantly (p<0.05) differentiated men with PCa from those with benign disease. Applying step-wise backward multivariate LR only total-PSA, % fPSA and sum-proPSA were retained and generated a receiver operator characteristic (ROC) curve with an area under the curve (AUC) of 77.25%. At 90% sensitivity, these three variables collectively achieved a specificity of 44% for the detection of PCa. Individually, the three retained variables had a specificity of 23% 33% and 15% respectively. CONCLUSIONS: Sum-pPSA, tPSA and % fPSA in combination significantly improves the specificity of early PCa detection in men with a total PSA of 4 to 10 ng/ml when compared with any of these individual PSA molecular forms measured alone. Source of Funding: Hybritech Inc., Division of Beckman Coulter.

1436 CLINICAL UTILITY OF PROPSA AND BPSA WHEN PERCENT FREE PSA IS BELOW 15% Alan W Partin", Leslie A Mangold, Lori J Sokoll, Daniel W Chan, Baltimore, MD; Stephen D Mikolajczyk, Harry J Linton, Cindy L Evans, Harry G Rittenhouse, San Diego, CA INTRODUCTION AND OBJECTIVE: Percent free PSA (25%) has greatly improved the specificity (20%) of PSA for early detection of prostate cancer while maintaining sensitivity (95%) when total PSA is between 4-10 ng/m!. Low percent free PSA values indicate higher risk of cancer yet only 30-50% of men with percent free PSA levels less than 15% have cancer at biopsy. The introduction of sub-forms of free PSA, proPSA and BPSA, has suggested a role for these new markers for early detection. Initial studies have demonstrated that proPSA improves detection when total PSA is between 2.5 and 4.0 ng/m!. In this study we investigated the clinical utility of proPSA and BPSA when percent free PSA is < 15%. METHODS: Archived sera from 161 consecutive men who were prospectively enrolled into our EDRN prostate cancer early detection biomarker program with percent free PSA <15% were studied (mean PSA 6.1 ng/ml, range 2-24 ng/ml). Mean age 62c!:.7 yrs. Diagnosis: Non-Cancer 59%, Cancer 41%. Total PSA, free PSA (Access Hybritech PSA, Beckman Coulter, Inc.), proPSA and BPSA (research use only, Beckman Coulter, Inc.) were measured for each sample. RESULTS: Overall AUC-ROC for the group for total and free-PSA were 0.51 and 0.54 respectively. There was no statistical difference between proPSA and free *Presenting author.

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PSA levels by diagnosis. BPSA and proPSAlBPSA both significantly improved cancer detection over percent free PSA (p<0.005). The AUC-ROC for proPSAI BPSA was 0.72 with a sensitivity and specificity of 90% and 46%, respectively, using a proPSAlBPSA cutoff of 0.61. CONCLUSIONS: These preliminary studies suggest that the ratio of proPSA (cancer PSA) and BPSA (benign PSA) can distinguish cancer with greater accuracy when percent free PSA is very low «15%). The measurement of proPSA and BPSA may therefore provide better clinical utility in this lower range of percent free PSA. Source of Funding: The Early Detection Research Network, NIHlNC1; Partial support: Beckman Coulter Inc.

1437 RATES OF CHANGE IN LEVELS OF HK2, FREE AND TOTAL PSA UP TO 20 YEARS BEFORE DIAGNOSIS OF PROSTATE CANCER David Ulmert*, Charlotte Becker, Malmo, Sweden; Thomas Bjork, Jan-Ake Nilsson, Malmo, Sweden; Goran Berglund, Hans Lilja, Malmo, Sweden INTRODUCTION AND OBJECTIVE: In a large population based study, we have reported that levels of all PSA-forms and hK2 in plasma are very strong predictors for prostate cancer (PCa) in samples collected up to 20 years before diagnosis of cancer. The purpose of this study was to investigate the rates of change and levels of PSA-forms and hK2 in a subset of these men who many years later were diagnosed with PCa. METHODS: Baseline samples were collected in 1974-86 from men (n=22,444; i.e. 75% acceptance rate) in Malmo, Sweden and born 1921, 19261942, 1944, 1946 and 1948-49. By Dec. 31st, 1999, 514 of these men had been diagnosed with PCa according to the Swedish Cancer Registry, while 215 of these men (with a median age of 48 years at baseline) had volunteered for a second blood-sampling 6 years after baseline. We analysed tPSA, fPSA, cPSA, percent fPSA, and hK2 in plasma from these men and 621 age-matched controls from the study population. HK2 was measured with an in-house research assay, total and free PSA with a commercial dual-label assay. RESULTS: Differences in levels of tPSA, cPSA, fPSA, percent fPSA, and hK2 in plasma were significantly different in cancer cases compared to controls (p
1438 A SINGLE NUCLEOTIDE POLYMORPHISM OF THE HUMAN KALLIKREIN-2 GENE (KLK2) HIGHLY CORRELATES WITH SERUM HK2 LEVELS AND IN COMBINATION ENHANCES PROSTATE CANCER DETECTION Robert K Nam*, William Zhang, John Trachtenberg, Eleftherios Diamandis, Michael A Jewett, Steven A Narod, Toronto, ON, Canada INTRODUCTION AND OBJECTIVE: To date, no genetic test is able to identify patients who are at high risk for prostate cancer in a clinical setting. We examined the relationship between a T for C substitution of the human kallikrein-2 gene (KLK2), circulating human kallikrein-2 (hK2) levels and prostate cancer risk. METHODS: We studied 1287 consecutive men who underwent prostate biopsies because of PSA level> 4 nglmL or an abnormal DRE. Serum and DNA samples were obtained prior to biopsy. Serum hK2 levels were measured using a sensitive immunofluorometric technique. The mutant and wild type alleles of the KLK2 gene were designated as the T allele and C allele, respectively. Unconditional logistic regression analysis was performed to estimate the odds ratio for having prostate cancer based on the KLK2 gene and levels, adjusting for total PSA, free:total PSA ratio, DRE, ethnicity, and family history. RESULTS: The mean PSA level was 12.3 ng/mL and the majority of the patients were white (84.0%). Of the 1287 men, 616 had cancer on any biopsy (cases) and 671 men had no cancer (controls). The overall distribution of the CC, CT and TT KLK2 genotypes was 55.1%, 38.2% and 6.8%, respectively. The

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median hK2 levels for men with the CC, CT and TT genotypes were 0.24, 0.18 and 0.062 ng/mL, respectively (p=O.OOOI). The adjusted odds ratios for having prostate cancer, for patients with TT and CT genotype compared to patients with CC genotype, were 3.93 (95% c.t. 1.8-8.5, p=0.005) and 1.67 (95% C.I.: 1.2-2.3, p=0.002), respectively. The adjusted odds ratios for having prostate cancer for patients in the fourth, third and second quartile levels of hK2 compared to patients in the first quartile were 2.31 (95% c.l.: 1.5-3.6, p=0.0002), 1.86 (95% C.l.: 1.2-2.9, p=0.005), and 1.04 (95% C.I.: 0.7-1.6, p=0.89), respectively. When we combined the KLK2 genotype with circulating hK2 levels, the adjusted odds ratios for having prostate cancer ranged from 1.87 (95% C.I.: 0.9-3.9, p=0.09) for patients with the lowest hK2 levels and the CC genotype to 13.92 (95% c.r: 6.6-29.2, p=O.OOOI) for patients with highest hK2 levels and at least one T allele. CONCLUSIONS: The KLK2 polymorphism and hK2 levels are correlated and, in combination, are highly predictive for prostate cancer. The hK2 test may help select patients to undergo repeat biopsy after an initial negative biopsy. Source of Funding: NCIC.

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Association between prostate cancer and SNP genotypes of BRG I nucleotide 33978 in exon 9 (SNP33978) was further analyzed in 48 prostate cancer patients and 36 benign controls. RESULTS: No somatic mutations were identified, however, five germline single nucleotide polymorphisms (SNPs) were revealed in CpG islands of the BRG I gene. Four of tbese SNPs were located in the coding sequences and one in an intronic region. Further association analysis of the BRG J SNP33978 showed that prostate cancer patients with TIT genotype were diagnosed at significantly younger ages than patients with TIC and CIC genotypes (p = 0.03). These young (age < 62 years) patients with the TIT genotype of SNP33978 were more frequently to have high grade, advanced stage and metastatic tumors than patients with the TIC or CIC genotype (p = 0.0134). CONCLUSIONS: Our results suggest that the TIT genotype of SNP33978 in the BRG I gene is associated with early onset of highly malignant prostate cancer. Source of Funding: None.

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ANALYSIS OF SERUM HUMAN KALLIKREIN 11 (HK11) FOR DISCRIMINATING BETWEEN PROSTATE CANCER AND BENIGN PROSTATIC HYPERPLASIA Terukazu Nakamura*,

LOW SERUM TESTOSTERONE LEVELS IN PROSTATE CANCER IS ASSOCIATED WITH A GENETIC RISK FACTOR

Toronto, ON, Canada; Andreas Scorilas, Athens, Greece; Carsten Stephan, Klaus lung, Berlin, Germany; Antoninus R Soosaipillai, Eleftherios P Diamandis, Toronto, ON, Canada INTRODUCTION AND OBJECTIVE: Prostate-specific antigen (PSA) is widely used as the most reliable tumor marker established so far. However, non-malignant prostatic diseases, especially benign prostatic hyperplasia (BPH) also cause serum PSA elevation, thus complicating the diagnosis of prostate cancer by PSA measurements alone. The determination of free PSA and its ratio to total PSA is now clinically established and is used to reduce the number of unnecessary prostate biopsies. Recently, we developed a specific immunoassay for human kallikreinll (hKII), one of the kallikrein gene family members, and found that hKII was highly expressed in prostate tissue. The aim of this study was to investigate if serum hK II levels could be used to discriminate prostate cancer from benign prostatic hyperplasia. METHODS: We analyzed for hKII, total PSA and % free PSA, a total of ISO serum samples from men with histologically confirmed benign prostatic hyperplasia (BPH; N = 64) or prostatic cancer (CaP; N = 86). Total and free PSA levels were measured by the Immulite PSA assay and hKII levels were measured by our previously published immunofluorometric assay. The analyses of differences between measured or calculated parameters, in the two groups, were performed with the non-parametric Mann-Whitney U test. Relationships between different variables were assessed by Spearman correlation coefficient. Receiver operating characteristic (ROC) curves were calculated for total PSA, % free PSA and hKIl/total PSA ratio and the area under the ROC curves (AUC) were analyzed by the Hanley and McNeil method. RESULTS: Serum hKII levels and the hKIl/total PSA ratio were both significantly lower in CaP patients than in BPH. In the subgroup of patients with % free PSA less than 20, an additional 54 % of BPH patients could have avoided biopsies using the hK II/total PSA ratio. ROC curve analysis demonstrated that the hKlll total PSA ratio (AUC = 0.81) and % free PSA (AUC =0.82) were much stronger predictors of prostate cancer than total PSA (AUC = 0.69). By combining % free PSA and hKIl/total PSA ratio, the number of unnecessary biopsies could be reduced by 80 %, in comparison to approximately 40 % by either %free PSA or hLII/ total PSA alone. CONCLUSIONS: Our preliminary data suggest that the hKIl/total PSA ratio could be a useful tumor marker for prostate cancer and could be combined with % free PSA to further significantly reduce the number of unnecessary prostatic biopsies. Source of Funding: None.

1440 THE Tff GENOTYPE OF SNP33978 IN THE BRGI GENE IS ASSOCIATED WITH EARLY ONSET OF HIGHLY MALIGNANT PROSTATE CANCER Alexander Valdman, Agneta Nordenskjoldd, Xiaolei Fang, Ayako Naito, Al-Shukri Salman, Catharina Larsson, Peter E Ekman, Chunde Li*, Stockholm, Sweden INTRODUCTION AND OBJECTIVE: Previous studies in hereditary and sporadic prostate cancer have indicated the existence of a tumor suppressor gene in chromosomal region 19p13. The BRG I gene in this region is one of the candidates, based on both the frequent inactivating mutations in human cancer cell lines, including the prostate cancer cell line DU145, and its functional properties. METHODS: To evaluate the possible involvement of BRGI gene in clinical prostate cancer, we carried out a complete mutation analysis of all 35 BRG I exons in tumor and constitutional DNA samples from 21 prostate cancer patients.

Georg Schatzl", Andrea Gsur, Stephan Madersbacher, Gerald Haidinger, Vienna, Austria; Michael Miksche, 1090, Austria; Michael Marberger, Vienna, Austria INTRODUCTION AND OBJECTIVE: Low serum testosterone levels and low CAG repeats are associated with high grade prostate cancer. Aim of this study was to examine the impact of four genetic polymorphisms within the androgen pathway on serum testosterone levels in prostate cancer patients. METHODS: In men with untreated bioptic verified prostate cancer an endocrine study such as testosterone (T), sex hormone binding globuline (SHBG), folJicle stimulating hormone (FSH), luteotropic hormone (LH), estradiol (E2), prostate volume and body mass index (BMI) were evaluated. Additionally, V89L and A49T polymorphisms of the type II steroid 5 alpha-reductase gene (SRD5A2), the polymorphic CAG repeats within exon I of the androgen receptor gene (AR) and cytochrom P 450c17alpha (CYP 17) were determined by polymerase chain reaction based methods using DNA from peripheral blood. RESULTS: A total of 101 men (65.4+1-7.2 yrs) were analysed. 34 men (33.7%) had a T level < 3 ng/ml. There was significant difference between the group T<3 ng/rnl compared to group T>3 ng/ml.regarding FSH (4.9+/-3.3 mIU/mL vs. 7.4+/-5.3 mIU/mL; p=0.016), E2 (19.7+1-9.2pg/mL vs. 24.8+111.2pg/mL; 0.026), calculated free testosterone (22.2+/-18.9 pg/mL vs. 65.0+/27.5 pg/mL; p7 had a higher risk (Odds ratio 2.33(IQR 0.5-10.83) to have T levels < 3 ng/ml. CONCLUSIONS: Low serum testosterone and low CAG repeats show significant association with high grade prostate cancer and could be an additional risk factor for poor outcome. Source of Funding: None.

1442 VARIANT OF CYP3A4 ASSOCIATED WITH RECURRENCE OF ANDROGEN INDEPENDENCE IN PROSTATE CANCER Mark B Fisher", Fernando 1 Bianco, Wael A Sakr, Richard Everson, Isaac 1 Powell, Detroit, MI INTRODUCTION AND OBJECTIVE: CYP3A4 is a cytochrome enzyme involved in the metabolism of testosterone. A genetic variant of CYP3A4 has been reported to be associated with a higher clinical grade and stage of prostate cancer (PCA). The heterozygous and homozygous alJele has been reported to be significantly more prevalent among African-American men. The purpose of this study is to examine the survival of men who have undergone radical prostatectomy to evaluate whether CYP3A4 gene variant is associated with disease progression. METHODS: 564 consecutative PCA patients with specimens available between 1991-1995 with RRP by academic staff were evaluated for CYP3A4 genotype (GG, GA or AG, AA) and variants. The genotype was correlated with standard clinicopathologic variables including age, race, PSA, pathological stage, and specimen Gleason score (7). Chi square was used for analysis. RESULTS: Median age 63.5 (40-77), median follow up 66 months (1-137), 92 % of Caucasians vs. 14% of African-Americans had the AA genotype. No correlation between genotype and Gleason score was observed. Table shows genotype expression of CYP3A4 and outcome correlation. CONCLUSIONS: This provocative result seems to suggest that the GG genotype carries a more aggressive course particularly in African-American males where 46% vs. I % of white males were identified with this genotype.

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Genotype Expression ofCYP3A4 and Outcome Correlation CYP3A4 GG GAorAG AA Pvalue

BioFail 38%

Androgen Independence 10%

~%

7%

W%

3%

0,001

0,02

Source of Funding: NIHJNCI grants CA8l240, CA84989, and a VIrtual Discovery Grant from the Karmanos Cancer Institute at Wayne State University.

1443 ANALYSIS OF SERUM PROTEOMIC PROFILES WITH SELDITOF AND ARTIFICIAL INTELLIGENCE BASED PATTERN RECOGNITION CAN HELP DETERMINE THE NEED FOR PROSTATE BIOPSY David K Ornstein", Chapel Hill, NC; Vincent Fusaro, Sally Ross, Ben A Hitt, Bethesda, MD; Gayle 1 Grigson, Henry A Bell, Raj S Pruthi, Chapel Hill, NC; Lance A Liotta, Emanuel F Petricoin, Bethesda, MD INTRODUCTION AND OBJECTIVE: Artificial intelligence based pattern recognition algorithms have been developed and successfully used to analyze complex serum proteomic data streams generated by surface enhanced laser desorption ionization time-of-f1ight mass spectroscopy (SELDI-TOF). We have previously shown that analysis of serum protein profiles by this novel approach can discriminate men with prostate cancer (CaP) from those with benign prostates (Petricoin et. al., INCI 2002). In this current study we demonstrate that SELDITOF analysis of serum proteins can be used to determine the need for prostate biopsy among men with PSA levels of 2.5 - 15 ng/ml or suspicious DRE. METHODS: Serum samples were collected from 114 men (108 with PSA 2.5 - 15 ng/ml and 6 with PSA < 2.5 ng/ml and suspicious DRE) prior to TRUSlBx, and serum profiles generated with the SELDI apparatus from Ciphergen Biosystems by applying 1 ul of serum to a hydrophobic interaction protein chip array. Profiles representing molecular masses of 1 - 20 kd were achieved by using alpha-cyano-4-hydroxy-cinnaminic acid for the matrix. The profiles were analyzed by KDE a pattern recognition algorithm developed by Correlogic Systems. Serum samples from 44 randomly selected men were used to "train" the diagnostic algorithm and the remaining 70 samples were analyzed in a blinded fashion. RESULTS: Of the 70 men used for the blinded analysis, 18 had CaP detected on initial biopsy. All 18 men with CaP were correctly identified by our proteomic analysis while 34 (68%) of the 52 men with benign biopsies were correctly classified. 12 of the 34 men whose biopsies were initially benign underwent repeat biopsy of which 4 were positive. 2 of these 4 men with CaP were correctly identified. Thus if SELDI-TOF had been used to determine the need for prostate biopsy, 68% of negative biopsies would have been eliminated but 10% of CaP would have been missed. CONCLUSIONS: Artificial intelligence based pattern recognition analysis of serum protein profiles generated by SELDI-TOF may be a useful diagnostic strategy that reduces "unnecessary" prostate biopsies without missing potentially harmful prostate cancers. Source of Funding: None.

1444 ASSOCIATION OF PREOPERATIVE PLASMA LEVELS OF VASCULAR ENDOTHELIAL GROWTH FACTOR AND SOLUBLE VASCULAR CELL ADHESION MARKER-! WITH BIOCHEMICAL PROGRESSION AFTER RADICAL PROSTATECTOMY Veronica A Anwuri*, Houston, TX.; Shahrokh F Shariat, Dallas, TX.; Dolores J Lamb, Houston, TX.; Michael W Kattan, New York, NY; Thomas M Wheeler, Kevin M Slawin, Houston, TX. INTRODUCTION AND OBJECTIVE: Angiogenesis has been shown to be a key process that enables tumor growth and metastasis in a wide variety of malignancies. Local expression of vascular endothelial growth factor (VEGF) and soluble vascular cell adhesion molecule-l (VCAM-l), critical factors that promote angiogenesis, are elevated in the circulation of patients with breast and colorectal carcinomas. We tested the hypothesis that elevated circulating levels of VEGF and soluble VCAM-l would be associated with prostate cancer progression and metastasis. METHODS: Levels of VEGF and sVCAM-l were measured in frozen plasma sample, obtained pre-operatively in 215 consecutive patients undergoing radical prostatectomy for clinically localized disease and nine men with untreated prostate cancer metastatic to bones using commercial ELISA assays (R&D Systems, Minneapolis, MN). RESULTS: Plasma VEGF and sVCAM-l levels were highest in patients with bone metastases (P<.OOl). Within the group of prostatectomy patients, while pre-operative plasma VEGF and sVCAM-llevels were elevated in patients with metastases to regional lymph nodes (P values <.001), only higher VEGF levels were associated higher biopsy and final Gleason sum (P=0.036 and P=0.040, *Presenting author.

respectively) and extraprostatic extension (P=0.047). Higher pre-operative VEGF levels was associated with lymph node involvement and biochemical progression (P=.043 and P=.020, respectively), when adjusted for the effects of standard pre-operative features. CONCLUSIONS: Plasma VEGF and soluble VCAM-l levels are markedly elevated in men with metastatic prostate cancer. Furthermore, both are independent predictors of biochemical progression after radical prostatectomy, presumably due to an association with microscopic metastatic disease already present at the time of surgery. After further validation studies, these markers may be candidate markers to enhance the performance of predictive normograms for prostate cancer. Source of Funding: Supported in part by Grant #IRG 93-034-06 from the American Cancer Society; the Austrian Science Fund; the Frost Foundation; Inc. and the National Cancer Institute Specialized Program of Research Excellence (SPORE CA58203).

Innovations in Minimally Invasive Approaches to the Genitourinary Tract Video Session Tuesday, April 29, 2003

1:00-3:00 PM

V1445 LAPAROSCOPIC TRAINING IN UROLOGY Ran Katz*, Andras Hoznek, Patrick Antiphon, Laurent Salomon, Alexandre De La Taille, Tomasz Borkowski, Dominique Chopin, Clement-Claude Abbou, CRETEIL, France INTRODUCTION AND OBJECTIVE: The role of laparoscopic surgery has increased during the last few years. The growing demand for laparoscopists in the urological field and the long learning curve lead to the development of various training modalities. METHODS: We developed a stepwise program to improve laparoscopic skills and teach complicated laparoscopic tasks. RESULTS: Pelvitrainers are inexpensive and readily available. The main goal of trainers is to develop hand and eye coordination in a two dimensional environment and familiarization with laparoscopic tools. Suturing and knot tying can be endlessly repeated. Models reproducing the spatial disposition of trocars and human topography allow to simulate reconstructive steps of specific procedures like vesicourethral anastomosis during laparoscopic radical prostatectomy. The main disadvantage of trainers is the impossiblilty to modalize dissection techniques on live tissues. Animal models allow practicing access creation to the peritoneal cavity or retroperitoneum, proper insufftation techniques and performing comlete procedures. The principles of dissection and hemostasis can be practiced. Several interventions can be performed including neophrectomy, pyeloplasty, vesicourethral anastomosis, colposuspension. However, animal antomy is quite different from the humen one. Cadavers have been used for several centuries in medicine to teach human anatomy. Although hemostasis can not be practiced on cadavers, this model allows to perfom entire procedures in condtions near to real clinical cases. CONCLUSIONS: The three presented training modalities allow to become familiar with all principal aspects of laparoscopic surgery. Recently, the introduction of robotic surgery opened the way to telesurgery and telementoring. With rapid development of computer technology, virtual reality will allow to elaborate real time interactive surgical simulators. Source of Funding: None.

V1446 EXTRAPERITONEAL LAPAROSCOPIC LYMPH NODE DISSECTION FOR EARLY STAGE TESTIS CANCER Akihiro Ito", Makoto Satoh, Koji Mitsuzuka, Seiichi Saito, Yoichi Arai, Sendai, Japan INTRODUCTION AND OBJECTIVE: We performed laparoscopic retroperitoneal lymph node dissection (RPLND) to assess the pathological status of retroperitoneal lymph nodes in Stage I disease or Stage IIa disease after chemotherapy. In this video, we present our procedure of extraperitoneal laparoscopic RPLND for Stage IIa non-seminomatous left-sided testis cancer after chemotherapy. METHODS: Under general anesthesia, the patient was placed in the supine position. A small incision was made out the McBurney's point" a third lateral point on the line between navel and iliac crest".Two lateral trocars are inserted under endoscopic view with great attention to avoid injury of the peritoneum. The peritoneum was separated from psoas muscle and retracted antero-medially. The ureter was identified on the common iliac artery and lifted up together with peritoneum. The space between the common iliac artery and the peritoneum was separated with a blunt dissection, adding to the effect of carbon dioxide insufflation