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(10.6%) out of 85 patients with a negative DSNB developed inguinal metastases which were submitted to LND: eight had pN2 and one pN3 disease. In one out of nine undetected DSN (cNx), inguinal nodal metastases developed at two months as pN2 disease. Most probably, lack of visualization is due to tumor blockage of lymphatic vessels drainage and rerouting to a neo sentinel node. We had only 5 (7.4%) metastases among the groins of 68 patients which had had negative DSNB. Overall, we recorded 16/94 affected groins (17%), so that LND was spared in 83% (78/94) of patients. On the other hand, the FN rate was 10.6% (9/85), leading to a relatively good NPV of 82% (70/85), but to only fair sensitivity of 40% (6/15). The efficiency rate of the whole procedure (groins of 94 patients) is even lower, as we detected only 6 (37.5%) out of 16 metastases. We did not find major complications. Nonetheless, one patient developed an unusual intradermal metastatic spread, possibly due to a lymphatic drainage alteration after DNSB. Conclusion: DNSB is a safe procedure, with limited major complications which was able to spare LND in 83% of our patients. As false-positives are not actually possible, only NPV (82.3%) and sensitivity (40%) can be calculated. Moreover, the efficiency rate of the intention to diagnose is even lower, as it was only possible to detect 37.5% of metastatic nodes early. Small numbers, intrinsic to the rarity of the disease, do not allow one to draw definitive conclusions. A close follow-up is mandatory in cases of a lack of groin signal (cNx), as well as in cases of negative bilateral DSNB. Patients with penile SCC should be evaluated in high volume centers, in order to achieve greater accuracy in this procedure and the best chance of cure. 1 Leijte JAP et al: Reliability and safety of current dynamic sentinel node biopsy for penile carcinoma. Eur Urol 52: 170- 177, 2007. 2 Pizzocaro G et al: EAU penile cancer guidelines 2009. Eur Urol 57: 1002-1012, 2010. 34 DIFFERENTIAL EXPRESSION <strong>OF</strong> MATRIX ATTACHMENT REGION-BINDING PROTEINS IN HUMAN PROSTATIC CANCER CELL LINES Paola Barboro1 , Erica Repaci1 , Valentina Traverso1 , Cristina D’arrigo2 and Cecilia Balbi1 1Istituto Nazionale per la Ricerca sul Cancro, Genova, Italy; 2Istituto per lo Studio delle Macromolecole Sez. Genova, C.N.R., Italy Background: Abnormal nuclear organization and alterations in the amount and distribution of heterochromatin have always been recognized as hallmarks of human cancer. However, neither the exact cause of these modifications nor how the activity/silencing of thousands genes can be orchestrated are 1828 ANTICANCER RESEARCH 31: 1807-1956 (2011) currently known. In eukaryotes, the genome is compartmentalized into chromatin domains by attachment to a supporting structure termed the nuclear matrix (NM). The interactions of chromatin with the NM occur via AT-rich DNA sequences called matrix attachment regions (MARs). MARs exhibit several functions, including the organization of chromatin loops, the augmentation of transcription and the facilitation of replication; moreover, the attachment of the genome to the NM is a dynamic event that is cell type- or cell cycle-dependent (1). Several MAR-binding proteins have been identified, some are dramatically dysregulated in tumor cells and their expression is often significantly correlated with the more aggressive phenotype (2). Likewise, modifications of the interactions between NM proteins and MARs might be related to the large-scale chromatin organization observed in carcinogenesis. Research carried out in our and other laboratories seeking prostate cancer (PCa) markers with improved diagnostic and prognostic features has identified some NM proteins that are differentially expressed in PCa with respect to non-tumor tissue; a few of these proteins were significantly correlated with tumor aggressiveness and/or risk of biochemical progression (3). These findings prompted us to characterize the changes in the NM protein–MAR bond in PCa. Materials and Methods: In this study, using a proteomic approach along with two-dimensional Southwestern analysis, we characterized the NM protein–MAR bond in two prostatic cancer cell lines: the androgen-responsive LNCaP and the androgen-resistant PC3. For binding experiments, a highly repetitive bent DNA sequence of 370-bp (XmnI) containing a base unpairing region was used as a probe. The variation of the expression of some MAR proteins was determined by quantitative Western blot. Both the nucleoplasm distribution and the co-localization with DNA were evaluated by confocal laser scanning microscopy. Using transmission electron microscopy (TEM) the high-order structure of chromatin was examined in the two cell lines. Results: Several NM proteins extracted from both cell lines strongly bind to the XnmI sequence. It is possible to separate these proteins into two groups: I. MAR-binding proteins present in a large quantity also in other cellular types (MARPa, MARPb, hnRNPs, Lamin A, Lamin B, Matrin3) that may correspond to the proteins in contact with the constitutive MARs (i.e. the sequences demarcating permanent domain boundaries in all cell types). II. MAR-binding proteins corresponding to celltype specific factors (PARPs, SATB1) that interact with facultative MARs (i.e. the sequences cell type- and activityrelated and dependent on cell differentiation). In PC3 with respect to LNCaP cells, several changes in the ability of proteins to bind XnmI sequence were observed: the signal intensity of Matrin3, SATB1 and all basic hnRNPs decreased, showing that these proteins bind the DNA more weakly or are down-regulated in more aggressive cells. In contrast, MARPa, MARPb, PARP and its fragments increased. Confocal
Abstracts of the <strong>21st</strong> Annual Meeting of the Italian Society of Uro-Oncology (SIUrO), 22-24 June, 2011, Naples, Italy microscopy demonstrated that also the co-localization of PARP with DNA was significantly lower. These changes were concomitant with a different compartmentalization of chromatin in the two cell lines, as shown by TEM. Conclusion: Our study provides evidence that several MARbinding proteins undergo extensive changes in PC3 with respect to LNCaP cells, showing that the interactions between NM and the base of the chromatin loop are involved in cell differentiation. These proteins may turn out to be an important tool in the understanding of PCa carcinogenesis and may be novel targets of anticancer drugs. 1 Chattopadhyay S and Pavithra L: MARs and MARBPs: Key modulators of gene regulation and disease manifestation. In: Chromatin and Disease. Kundu TK and Dasgupta D (eds.). Springer, pp. 213-230. 2007. 2 Han HJ, Russo J, Kohwi Y and Kohwi-Shigematsu T: SATB1 reprogrammes gene expression to promote breast tumour growth and metastasis. Nature 452: 187-193, 2008. 3 Barboro P, Rubagotti A, Boccardo F, Carnemolla B, Darrigo C, Patrone E and Balbi C: Nuclear matrix protein expression in prostate cancer: possible prognostic and diagnostic applications. Anticancer Res 25: 3999-4004, 2005. 35 ROBOT-ASSISTED LAPAROSCOPIC RADICAL CYSTECTOMY AND INTRACORPOREAL URINARY DIVERSION Roberto Nucciotti, Valerio Pizzuti, Francesco Mengoni, Fabrizio Viggiani, Fabio Massimo Costantini, Giandomenico Passavanti and Alessandro Bragaglia U.O. Urologia Ospedale della Misericordia, Grosseto, Italy Aim: To describe our technique of robotic cystectomy and intracorporeal formation of Studer neobladder for treatment of carcinoma of the bladder. Materials and Methods: We describe our surgical technique, stepwise describing the surgical procedure and pathologic outcomes. After cystectomy, the reconstruction phase starts from the ileo-urethral anastomosis about 40 cm from the ileocecal valve. We then proceeded to section the ileum through endoGIA. Detubularization of ileum neobladder is carried out using cold robotic scissors. Studer reconfiguration is performed by pds running suture. To reduce urinary reflux, we prefer the Wallace type of anastomosis, with a pro-peristaltic segment of nondetubularized ileum. Results: We performed 15 robotic-assisted intracorporeal neobladder procedures. Mean operating-room time of all patients was 6.3 h and mean surgical blood loss was 430 ml. We prefer to stent the ureteral anastomosis for two weeks. We have recorded only one major postoperative complication: an acute compartment syndrome of the lower leg. Conclusion: The clinical and oncologic follow-up of patients undergoing robotassisted intracorporeal urinary diversion appears to be favorable in the short term. As our follow-up increases, we should expect to continue to truly define the long-term clinical appropriateness and oncologic success of this procedure. 36 PROSTATE CANCER BEFORE AND AFTER RENAL TRANSPLANTATION: OUR EXPERIENCE Fedele Lasaponara1 , Andrea Buffardi1 , Giovanni Pasquale1 , Omidreza Sedigh1 , Andrea Bosio1 , Paolo DeStefanis1 , Mariateresa Carchedi1 , Giuseppe Paolo Segoloni2 and Dario Fontana1 1S.C. Urologia 2 U. and 2S.C. Nefrologia Dialisi e Trapianto Renale U., A.O.U. San Giovanni Battista "Molinette", Turin, Italy Background: Prostate cancer is the most common genitourinary malignancy in male renal transplant recipients. Renal transplantation is the preferred therapy in patients with endstage kidney disease. Improvements in immunosuppressive therapy and in surgical techniques offer good renal allograft and patient survival times. Furthermore, the increased age of recipients leads to a high number of diagnoses of prostate cancer at the time of placement on the transplantation waiting list. Patients and Methods: From 1981 to 2010, 2,499 renal transplantations were performed at our center, 1610 of which were for males (64.4%). We found thirteen cases (Group 1) of prostate cancer at the time of placement on the waiting list and eight cases (Group 2) after kidney transplantation (from 1 month to 12 years after grafting). Patients in Group 1 underwent radical treatment: retropubic prostatectomy in three cases, perineal prostatectomy in six cases, HIFU in four cases (and one more HIFU after perineal prostatectomy for a pT3aNx with involved margins). After treatment, the follow-up for these patients consisted of a physical examination (including a DRE) and serial serum PSA measurements every three months. Patients in Group 2 underwent: perineal prostatectomy in three cases and radiation therapy in one case (for localized tumor), TUR-P in three cases (incidental tumors) followed by watchfulwaiting, hormonal therapy with LH-RH analogs in one case (a patient with a Gleason Score of 5 who refused radical therapies). After treatment, the follow-up consisted of a physical examination (including a DRE) and serial serum PSA measurements every three months, abdominal US and chest Xrays every six months. Results: Patients in Group 1 submitted only to radical surgery (eight cases) were placed on the waiting list six months after the procedure: the mean serum PSA level was 0.03 ng/ml and they were considered free from disease. At the time of writing, four of them have received grafts, one is dead from infective complications, and three are still on the 1829
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255 ACCURACY OF ENDORECTAL MRI AND
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Nucciotti R, 35 Oderda M, 37, 38, 1
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