Alterations in methylation of CpG dinucleotides at the 5 position of deoxycytidine residues (5mC) are a hallmark of cancer cells, including testicular germ cell tumors. Virtually all testicular germ cell tumors are believed to be derived from intratubular germ cell neoplasia unclassified (IGCNU), which is thought to arise from primordial germ cells. Prior studies revealed that seminomas contain reduced levels of global DNA methylation as compared with nonseminomatous germ cell tumors. Smiraglia et al have proposed a model whereby seminomas arise from IGCNU cells derived from primordial germ cells that have undergone 5mC erasure, and nonseminomas arise from IGCNU cells derived from primordial germ cells that have already undergone de novo methylation after the original erasure of methylation and contain normal 5mC levels. Yet the methylation status of IGCNU has not been determined previously. We used immunohistochemical staining against 5mC to evaluate global methylation in IGCNU and associated invasive testicular germ cell tumors. Strikingly, staining for 5mC was undetectable (or markedly reduced) in the majority of IGCNU and seminomas, yet there was robust staining in nonseminomatous germ cell tumors. The lack of staining for 5mC in IGCNU and seminomas was also found in mixed germ cell tumors containing both seminomatous and nonseminomatous components. Lack of 5mC staining was not related to a lack of the maintenance methyltransferase (DNA methyltransferase 1) protein. We conclude that testicular germ cell tumors are derived in most cases from IGCNU cells that have undergone developmentally programmed 5mC erasure and that the degree of subsequent de novo methylation is most closely related to the differentiation state of the neoplastic cells. That is, IGCNU cells and seminoma cells remain unmethylated, whereas all other histological types appear to arise after de novo methylation.

Tissues from patients diagnosed with prostate cancer and PIN exhibit, on average, locally increased SMO expression in regions of prostatic disease and higher overall SMO expression in prostatic epithelial cells compared to healthy individuals. Further studies are warranted to directly examine the role of SMO-produced ROS in prostate carcinogenesis.

Background Bladder urothelial carcinoma (UrCa) proclaims high rates of mortality and morbidity. Identifying novel molecular prognostic factors and targets of therapy is crucial. mTOR pathway plays a pivotal role in establishing cell shape, migration and proliferation. Design TMA were constructed from 132 cystectomies (1994-2002). IHC was performed for Pten, c-Myc, p27, phosAkt, phosS6, and 4E-BP1. Markers were evaluated for pattern, percent and intensity of staining. Results Mean length of F/U was 62.6 months (1-182). Disease progression, overall survival (OS) and disease specific survival (DSS) rates were 42%, 60% and 68%, respectively. Pten showed loss of expression in 35% of UrCa. All markers showed lower expression in invasive UrCa compared to benign urothelium with the exception of 4E-BP1. Pten, p27, phosAkt, phosS6 and 4E-BP1 expression correlated with pathologic stage (pT; p<0.03). Pten, 4E-BP1, and phosAkt expression correlated with divergent aggressive histology and invasion. PhosS6 expression inversely predicted OS (p=0.01), DSS (p=0.001) and progression (p=0.05). c-Myc expression inversely predicted progression (p=0.01). In a multivariate analysis model that included: TNM stage grouping, divergent aggressive histology, concomitant CIS, phosS6 and c-Myc expression; phosS6 was an independent predictor of DSS (p=0.03; HR: -.19) while c-Myc was an independent predictor of progression (p=0.02; HR:-.38). In a second model substituting organ confined disease and lymph node status for TNM stage grouping, phosS6 and c-Myc remained independent predictors of DSS (p=0.03; HR: -.21) and progression (p=0.03; HR:-.34), respectively. Conclusions We found an overall downregulation of mTOR pathway in UrCa. PhosS6 independently predicted DSS and c-Myc independently predicted progression.