C16 PREDICTING PROSTATE CANCER BY DETECTING GSTP1 ABERRANT PROMOTER METHYLATION IN SERUM DNA

Introduction & Objectives: Recent studies suggest that chronic inflammation is crucial in the development and progression of prostate cancer (CaP). Inflammatory response depends upon differential production of cytokines, which is correlated with single-nucleotide polymorphisms (SNPs) of cytokine genes. So far, the studies of cytokine genes’ SNPs have shown contradictory results and propose ethnical dependency of SNPs. Interleukin-10 (IL10) has dual properties: it can reduce tumor angiogenesis, but can also enable tumor cells to «escape» the immune response with its anti-inflammatory action. We here investigated the association of two IL10 SNPs (IL10-819 C>T and IL10-592 C>A) with CaP risk in Eastern Croatian population. Material & Methods: 120 CaP patients and 120 benign prostatic hyperplasia (BPH) controls treated at the Clinic for Urology, University Hospital Centre Osijek were included in this study, after giving informed consent. Groups were agematched. Two subgroups were formed according to Gleason score (≥4+3 and ≤3+4). To determine the IL10 SNPs, genomic DNA was extracted from 200 µl ethylenediaminetetraacetic acid (EDTA) whole blood using commercially available kits. IL10 gene polymorphism was assessed by real-time PCR method with melting curve analysis. Descriptive statistical analysis was performed with SPSS 19.0 statistical program (SPSS Inc., Chicago, Ill, USA); x2 test was used to determine the differences between the groups and Kendall’s tau-b test for linear-by-linear association. Results: For the IL10-819 and -592 SNPs there were no significant differences between cases and controls. The -819 TT variant was associated with a 2-fold decreased risk for CaP, but the trend was not significant (OR = 0.5, 95% CI 0.161.54, p trend = 0.6). The -592 allele distribution was identical in cases and controls. When comparing the subgroups, the -819 TT variant showed a slightly increased risk for higher scores (OR = 1.23, 95% CI 0.19-7.84, p trend = 0.88). The two IL10 SNPs were also tested according to PSA levels (<10 and ≥10; >20 and ≥20) but again the groups did not differ. Conclusions: Although there are studies that report the association between the IL10 SNPs and CaP risk, recent meta-analysis of 10 studies on the subject found no significant association. Our analysis also shows lack of such an association for Eastern Croatian Population. These findings, taken together with contradictory results of other cytokine genes’ SNPs imply the ethnical dependency of such polymorphisms. Also, combinations and interactions of SNPs might have a greater impact on CaP risk than just one SNP. Future studies will define true importance of cytokine genes’ SNPs in the development and progression of CaP.

C13

Analyzing the lymphatic drainage of prostate in patients with prostate cancer based on lymphoscintigraphy

Klijer R., Muc K., Bar K., Urban M., Buraczynski P. Medical Univeristy, Dept. of Urology and Uro-Oncology of Medical University, Lublin, Poland Introduction & Objectives: Lymph node evaluation is an important element of correct diagnosis and treatment of prostate cancer. Great deal of analysis shows that lymphatic drainage of prostate gland is multidirectional. In many cases minimal dissection consisted of obturator fossa lymph nodes removal is routine procedure in the treatment of locally advanced prostate cancer. Aim of this analysis is to evaluate lymphatic drainage in locally advanced prostate cancer. Material & Methods: During years 2005-2010 59 patients before radical prostatectomy underwent lymphoscintigraphy in the Department of Urology and Uro-oncology of Medical University in Lublin. Radioactive tracer (99mTc Nanocoll) in a volume of 1 ml and 100MBq activity (2.7 mCi) was administered centrally to each lobe of the prostate under control of transrectal ultrasonographic head in about 24 hours before surgery. Lymphoscintigraphy in antero-posterior projection was performed about two hours after administration of the tracer. We analyzed Lymph flow to particular groups of lymph nodes (such as obturator fossa lymph nodes, external iliac lymph nodes, internal iliac lymph nodes, common iliac lymph nodes, aortic lymph nodes) and investigated its relationship with the clinical stage. Results: The study included 59 patients aged from 47 to 74 years, a mean age 65.5. Lymph drainage was visualized in all patients, unilaterally – in 27 cases (46%) and bilaterally – in 32 cases (54%). 34 patients (58%) were in stage pT2c, 13 (22%) in pT2b, 3 (5.1%) in pT2a, 7 (11.9%) in pT3b, 2 (3.4%) in pT3a. Preoperative PSA ranged from 2.7 ng/ml to 34.20 ng/ml, middling 6.6 ng/ml. Lymph flow was as follows: external iliac lymph nodes manifested in 38 cases (64%), obturator fossa lymph nodes – in 16 cases (27%), internal iliac lymph nodes – in 15 cases (25%), paraaortic – in 1 case, common iliac lymph nodes – in 1 case. Conclusions: On the basis of lymphoscintigraphy in prostate cancer lymph generally flows to external iliac lymph nodes. Rarely lymph flows to the obturator fossa lymph nodes and internal iliac lymph nodes. There was no correlation between the ways of lymph drainage and clinical stage.

C14

Relationship between obesity and prostate cancer

Cosic I.1, Sudarevic B.1, Mandic S.2, Horvat V.2, Marczi S.3, Simunovic D.1, Galic J.1 1 University Hospital Center Osijek, Dept. of Urology, Osijek, Croatia, 2University Hospital Center Osijek, Dept. of Clinical Biochemistry, Osijek, Croatia, 3University

Eur Urol Suppl 2011;10(9):616

Hospital Center Osijek, Dept. of Molecular Biology, Osijek, Croatia Introduction & Objectives: It is well known that, beside age, race and familiar factor, dietary factors play big role in epidemiology of prostate cancer (CaP). The Western lifestyle, particularly the higher intake of fat, meat, and dairy products, is also associated with risk for CaP. Obesity is also linked to CaP, mainly with mortality rather than incidence. We wanted to see and show if there is difference in body mass index (BMI) in patients with biopsy diagnosed CaP and patients in control group. Material & Methods: We have included 235 patients in our research was (age 42-84 yrs.). In all patients transrectal biopsy of prostate was done. In 117 patients out of 235 is diagnosed CaP, in rest of 118 patients patohistological finding did not show CaP (control group). BMI and PSA level were determinated to all patients. Student t-test was used to assign difference between BMI in our 2 groups. Results: Mean BMI in group of patients with CaP was 27.625 kg/m2 (SD 5.680), mean BMI in control group was 27.692 kg/m2 (SD 3.987). There was no statistical significant difference in BMI in out two groups. When we grouped our patients according to their BMI (underweight < 18,5 kg/m2; normal 18,5-25 kg/m2; overweight 25-30 kg/m2 and obese >30 kg/m2), we noticed decrease of PSA level with increase of BMI. In our research there was no underweight patients, 64 of them were with normal BMI and mean PSA 69.48 ng/ml; 120 of them were overweight with mean PSA 55.15 ng/ml and 51 of them were obese patients with mean PSA 33.16 ng/ml. Conclusions: General adiposity, as measured by BMI, is not in relationship with CaP incidention in our research, but obese is connected with lower PSA level with possible hemodilution in background. Question is – should we decrease the threshold of PSA in obese patients for prostate biopsy?

C15

Expression of TopBP1 gene in normal and cancerous prostate tissue

Wilkosz J.1, Różanski W.1, Markowski M.1, Lipinski M.1, Forma E.2, Bryś M.2 1 Medical University of Lodz, Dept. of Urology, Lodz, Poland, 2University of Lodz, Dept. of Cytobiochemistry, Lodz, Poland Introduction & Objectives: TopBP1 (topoisomerase II β binding protein1) is one of the proteins involved in DNA damage response. the most striking feature of TopBP1 is that it has eight BRCT (BRCA1 C-terminal) domains. These were first identified in BRACA1 (breast cancer gene 1) and have since been detected in a host of cellular proteins. Abberant of TopBP1 expression may be involved in the deregulation of DNA damage response, cell cycle checkpoints and can have pathological cosequences. Therefore, the current study was aimed at investigating TopBP1 gene expression in normal and cancerous prostate tissue. Material & Methods: Real-time quantitave PCR was used to analysis of TopBP1 expression in 33 normal prostate gland, 40 benign prostatic hyperplasia (BPH), 9 prostatic intraepithelial neoplasia (PIN), and 73 prostate cancer samples. the cancer sprcimens were assessed regarding they tumor stage according to TNM classification (24 tumors were stage T1, 26 stage T2, 5 stage T3 and 18 stage T4) and Gleason scores (45 samples with Gleason score <=7 and 28 samples with Gleason score>7). All tumors were classified as N0 and M0. Results: The expression of TopBP1 gene in different normal and pathological tissues was estimated at the mRNA level and normalized to GAPDH expression level. Positive expression of TopBP1 was found in 97.0% of normal prostate gland and was more frequently observed compare to prostate cancer (76.7% p=0.04). level of TopBP1 mRNA was significant higher in normal tissue than in prostate cancer samples(p=0.006). the expression of TopBP1 mRNA was significantly lower in stage T3 and T4 tumors than in stage T1 (p=0.04 and 0.005 respectively). There was no statistically significant difference in TopBP1 expression in association with Gleason scores (p=0.06). in case of BPH and PIN, level of TopBP1 mRNA was lowe compare to normal prostate tissue but these differences were not statistically significant. Conclusions: Here we show deregulation of TopBP1 expression in studied normal and pathological prostate tissues. the findings in this study suggest that TopBP1 may be related to neoplastic transformation of prostate and may become important for tumor progression.

C16

Predicting prostate cancer by detecting GSTP1 aberrant promoter methylation in serum DNA

Dumache R.1, Negru S.2, David D.L.1, Kaycsa A.1, Puiu M.3, Ionescu D.4 1 University of Medicine and Pharmacy ‘’Victor Babes’’, Dept. of Biochemistry, Timisoara, Romania, 2University of Medicine and Pharmacy ‘’Victor Babes’’, Dept. of Oncology, Timisoara, Romania, 3University of Medicine and Pharmacy ‘’Victor Babes’’, Dept. of Medical Genetics, Timisoara, Romania, 4University of Medicine and Pharmacy ‘’Victor Babes’’, Dept. of Toxicology, Timisoara, Romania Introduction & Objectives: Hypermethylation of CpG islands in promoter regions is recognized as an important early event in prostate carcinogenesis and, detection of methylated DNA is considered a potential biomarker in early detection of PCa.Hypermethylation of regulatory sequences at the glutathione S-transferase pi (GSTP1) gene locus is found in the majority (>90%) of primary

prostate carcinomas, but not in normal prostatic tissue or other normal tissues. The aim of our study was to explore the diagnostic value of detecting aberrant promoter hypermethylation of GSTP1 gene in serum samples from patients with the diagnostic of prostate adenocarcinoma, as a noninvasive epigenetic biomarker for early molecular detection of PCa. Material & Methods: For our study we collected serum samples from 34 patients with the histologically confirmed cases of prostate adenocarcinoma, Gleason score in a range of 3 to 9, and serum PSA was in a range of 4 ng/ml to 28 ng/ml, and 54 cases with BPH. Patients with BPH were used as control subjects. Genomic DNA was extracted from serum samples using Zymo Research Serum DNA kit™. To evaluate the methylation status of the GSTP1 gene we used the methylationspecific PCR (MSP) method. Results: By MSP method the GSTP1 promoter hypermethylation was detected in 32 from 34 PCa samples (94.12%), whereas none of the BPH samples showed aberrant methylation. Conclusions: GSTP1 promoter hypermethylation distinguishes between PCa and BPH, and thus it could be used as an epigenetic biomarker for PCa screening and early molecular detection of this disease.

C17

Prostate targeted biopsy using cadencecontrast pulse sequencing technology

Jinga V.1, Budau M.2, Braticevici B.1, Radavoi G.D.1, Calin C.1, Badescu D.1, Diaconescu D.1, Onu M.2 1 “Th. Burghele Hospital”, Dept. of Urology, Bucharest, Romania, 2“Th. Burghele Hospital”, Dept. of Radiology, Bucharest, Romania Introduction & Objectives: Transrectal ultrasound-guided needle biopsy of the prostate may not reveal an important amount of clinically relevant cancers. Cancer tissue shows an increased in microvessel density and abnormal pattern of vascularity. Ultrasound contrast agents allow for a more complete delineation of the neovascular anatomy by enhancing the signal strength from small vessels. Cadence contrast pulse sequencing technology (CPS) is using a low mechanical index which protects the microbubbles and increase the parenchimal enhancement provided by ultrasonographic contrast agent. The aim of this study is to compare the prostate cancer detection rate of CPS-targeted biopsies with conventional ultrasound guided systematic biopsies. Material & Methods: A prospective study was performed on 54 men, mean age 62,5 (range 48-75), referred for prostate biopsy due to an increased PSA or abnormal DRE. During the administration of the ultrasound contrast agent (SonoVue, Bracco - 2.4 ml i.v. bolus injection, with a maximum dose of 4.8 mL), a prostate scanning was performed, from base to the apex, to assess suspicious enhancing areas of the peripheral zone. We have seen a rapid and increased contrast enhancement in only 33 patients (61,11%) and these patients were subjected to CPS-guided targeted biopsies (5 or less cores). Subsequently a 10 core systematic biopsy was taken in all 54 men. Results: CPS-targeted biopsy revealed cancer in 20 of 33 patients (60,60%) and overal detection rate using targeted biopsies was 37,03% (20/54). Systematic biopsy detected cancer in 17 of the 54 patients (31,48%). Sensitivity in PCa detection was 86,9% for CPS-targeted biopsy (20/23) and 73,91% (17/23) for systematic biopsy. Cancer was detected by targeted biopsy alone in 6 patients and by systematic biopsy alone in 3 patients. CPS-targeted cores were positive in 44 of 114 cores (38,59%) and in 42 of 540 (7,77%) systematic biopsy cores. Conclusions: 1. CPS-targeted biopsy are superior to systematic prostate biopsy in CaP detection. The targeted approach detect more cancers with a lower number of biopsy cores. 2. An increase in cancer detection was achieved by combining targeted and systematic techniques3. CPS-technique allow for the detection of patients with a very high likelihood of having prostate cancer

C18

Relationship between the positive core percentage and Gleason score with perineural invasion in patient with prostate cancer on prostate biopsy

Gorgel S.N.1, Ozdamar Y.K.2, Kara C.1, Ozer K.1 1 Izmir Ataturk Training and Research Hospital, Dept. of 1st Urology Clinic, Izmir, Turkey, 2Manisa Merkez Efendi State Hospital, Dept. of Urology, Manisa, Turkey Introduction & Objectives: We investigated relationship between the positive core percentage and gleason score with perineural invasion (PNI) in patients who underwent transrectal ultrasonography (TRUSG) guided prostate biopsy because of high prostate specific antigen (PSA) values and abnormal digital rectal examination. Material & Methods: A total of 1135 patients underwent 10-quadrant TRUSG guide prostate biopsy because of high PSA values and abnormal digital rectal examination in our clinic between 2004-2011. Prostate adenocarcinoma was detected in 274 patients (24.14%). Perineural invasion was demonstrated in 46 (16.78%) of prostate carcinoma cases. Cases with and without PNI were compared in terms of age, total PSA, prostate volume, PSA density, gleason score, positive core percentage and rectal examination. Patients were divided into 2 groups of total gleason score ≤6 and total gleason score >6, also positive core percentages

were divided into 2 groups (≤33% and > 33%) and these groups were compared in terms of perineural invasion. Results: The mean age was 70.29±7.98 years in patients with perineural invasion and 69.95±8.68 in patients without perineural invasion (p=0.095). The mean PSA level was 15.95±14.68 mg/dl in patients with perineural invasion and 38.72 ± 214 in patients without perineural invasion (p=0.473). There was no statistically singnificant difference in terms of age, total PSA, prostate volume, PSA density and rectal examination. However gleason score and positive core percentage were significantly higher in the patient with perineural invasion (respectively p<0.001 and p<0.001). Perineural invasion was significantly high in gleason score >6 and positive core percentage > 33% groups (respectively p<0.001 and p<0.001). Conclusions: Gleason score and positive cor percentage are indicators of poor prognosis in patients with prostate cancer. In this study, we observed a strong correlation between the gleason score and positif core percentage with perineural invasion. We believe that perineural invasion is an important prognostic factor to determine treatment options in patients with prostate cancer.

C19

Immunohistochemical expression of caveolin-1 on biopsy cores in detection of significant prostate cancer in low risk patients

Tomaskovic I.1, Tomic M.1, Ulamec M.2, Kruslin B.2, Grubisic I.1, Justinic D.1, Ruzic B.1, Spajic B.1, Reljic A.1, Trnski D.1 1 University Hospital „Sestre Milosrdnice“, Dept. of Urology, Zagreb, Croatia, 2 University Hospital „Sestre Milosrdnice“, Dept. of Pathology, Zagreb, Croatia Introduction & Objectives: In this retrospective non-randomized study we examined utility of imunohistochemical expression o caveolin-1 protein on preoperative biopsy cores in detection of significant prostate cancer in low risk patients. The existing preoperative criteria for differentiation between significant and insignificant prostate cancer in low risk patients appear to be inadequate. Material & Methods: 873 patients who underwent radical prostatectomy from 01.01.1998. till 31.12.2007. in our institution were included in the study. 105 patients fulfilled the most stringent preoperative criteria for insignificant protate cancer (PSA ≤ 10 ng/ml, Gleason sum ≤6, ≤33% positive biopsy cores, ≤50% of tumor in a core). Archival paraffin wax embedded prostatic biopsy cores were immunostained for caveolin-1 protein. Preoperative clinicopathological parameters and IHC expression caveolin-1 on tumor tissue in biopsy cores were correlated with tumor significance on postprostatectomy surgical specimens. Insignificant tumor was considered if <5% of final specimen contained tumor and Gleason sum was ≤6. Results: Postoperatively, 49 (46,7%) patients had insignificant and 56 (53.3%) had significant prostate cancer. Mann-Whitney non parametric test showed statisically significant difference in epithelial (p <0,001) and stromal (p =0,001) IHC expression of caveolin-1 between two groups of patients (insignificant vs. significant). Fisher´s exact test showed statisically significant difference between the groups in prostate volume (p=0.030), PSA (p=0,014), PSAD (p=0,014), cancer volume (p<0,001). No difference was found in age (p=0.255), number of biopsy cores (p=0,934) and tumor percentage in the core (p=0.086) between two groups. Conclusions: Epithelial expression of caveolin-1 appeared to be a risk factor for significant prostate cancer in low risk patients. Its´ expression on preoperative biopsy cores might be used as an additional criterion to existing ones in identification of significant prostate cancer in the group of low risk patients. This may influence the selection of therapeutic approach to low risk prostate cancer patients.

C20

Prostatic lesions with inverted pattern

Dema A.1, Taban S.2, Borda A.3 1 Victor Babes University of Medicine and Pharmacy, Dept. of Pathology, Timisoara, Romania, 2Clinical Emergency County Hospital, Dept. of Pathology, Timisoara, Romania, 3University of Medicine and Pharmacy, Dept. of Pathology, Targu Mures, Romania Introduction & Objectives: The inverted pattern, with the polarization change of the secretor cells nuclei, is a very rare finding in prostate lesions, being especially associated with high-grade prostate intraepithelial neoplasia (HG-PIN) foci. This paper presents the morphological aspects, along some immunohistochemisty (IHC) aspects, of the prostate lesions with inverted pattern (hobnail). Material & Methods: We have examined 2,000 consecutive prostatic specimens and we have selected all the prostate lesions with inverted pattern. Additional sections of paraffin blocks for these lesions were immunohistochemically marked with anti-basal cells antibodies (p63 and HMWCK), and with anti-alpha-methylacylCoA-racemase (AMACR), EnVision System, DAB. Results: We have observed 5 cases of prostatic specimens, in which the nuclei of the principal cells were, at least focally, oriented to the glandular lumen. The 5 cases were of patients aged between 65 and 78 years. One case was interpreted, based on cytological atypia and on the association with the classical patterns inside the same gland, as inverted HG-PIN associated with poorly differentiated carcinoma, on a transurethral resection (TUR) specimen. In the other 4 cases the architectural modification was detected on TUR fragments (3 cases), in association

Eur Urol Suppl 2011;10(9):617