Objective: DNA damage repair mechanisms are a source of genetic mutation and are believed to play an important role in human cancer. Human 8-oxoguanine DNA glycosylase I (hOGG1) is involved in the recognition and repair of DNA damage. The value of the hOGG1 genotype as a prognostic indicator for bladder cancer (BC) was assessed using a novel technological approach.Materials and methods: The association between genetic polymorphism of hOGG1 codon 326 and clinicopathologic characteristics of 337 patients with BC was analyzed using peptide nucleic acid (PNA)-mediated real-time PCR clamping.Results: Tumor grade and size were significantly associated with the hOGG1 codon 326 genotype in non-muscle-invasive bladder cancer (NMIBC). The Cys326Cys polymorphism was significantly associated with progression and cancer specific survival in patients with muscle-invasive bladder cancer (MIBC). Multivariate Cox regression analysis indicated that the hOGG1 Cys326Cys polymorphism is associated with a protective effect on progression and a more dominant survival benefit than the Ser326Ser polymorphism in MIBC (hazard ratio 0.284 and 0.305, respectively).Conclusions: Analysis of genotypes and clinical data for 337 BC patients indicates that the hOGG1 genotype may be a useful prognostic genetic marker for MIBC. Crown Copyright (C) 2012 Published by Elsevier Inc. All rights reserved.

Purpose: Abnormal DNA methylation is associated with many human cancers. The aim of the present study was to identify novel methylation markers in prostate cancer (PCa) by microarray analysis and to test whether these markers could discriminate normal and PCa cells. Experimental Design: Microarray-based DNA methylation and gene expression profiling was carried out using a panel of PCa cell lines and a control normal prostate cell line. The methylation status of candidate genes in prostate cell lines was confirmed by real-time reverse transcriptase-PCR, bisulfite sequencing analysis, and treatment with a demethylation agent. DNA methylation and gene expression analysis in 203 human prostate specimens, including 106 PCa and 97 benign prostate hyperplasia (BPH), were carried out. Further validation using microarray gene expression data from the Gene Expression Omnibus (GEO) was carried out. Results: Epidermal growth factor–containing fibulin-like extracellular matrix protein 1 (EFEMP1) was identified as a lead candidate methylation marker for PCa. The gene expression level of EFEMP1 was significantly higher in tissue samples from patients with BPH than in those with PCa (P < 0.001). The sensitivity and specificity of EFEMP1 methylation status in discriminating between PCa and BPH reached 95.3% (101 of 106) and 86.6% (84 of 97), respectively. From the GEO data set, we confirmed that the expression level of EFEMP1 was significantly different between PCa and BPH. Conclusion: Genome-wide characterization of DNA methylation profiles enabled the identification of EFEMP1 aberrant methylation patterns in PCa. EFEMP1 might be a useful indicator for the detection of PCa. Clin Cancer Res; 17(13); 4523–30. ©2011 AACR.

Background While several molecular markers of bladder cancer prognosis have been identified, the limited value of current prognostic markers has created the need for new molecular indicators of bladder cancer outcomes. The aim of this study was to identify genetic signatures associated with disease prognosis in bladder cancer. Results We used 272 primary bladder cancer specimens for microarray analysis and real-time reverse transcriptase polymerase chain reaction (RT-PCR) analysis. Microarray gene expression analysis of randomly selected 165 primary bladder cancer specimens as an original cohort was carried out. Risk scores were applied to stratify prognosis-related gene classifiers. Prognosis-related gene classifiers were individually analyzed with tumor invasiveness (non-muscle invasive bladder cancer [NMIBC] and muscle invasive bladder cancer [MIBC]) and prognosis. We validated selected gene classifiers using RT-PCR in the original (165) and independent (107) cohorts. Ninety-seven genes related to disease progression among NMIBC patients were identified by microarray data analysis. Eight genes, a progression-related gene classifier in NMIBC, were selected for RT-PCR. The progression-related gene classifier in patients with NMIBC was closely correlated with progression in both original and independent cohorts. Furthermore, no patient with NMIBC in the good-prognosis signature group experienced cancer progression. Conclusions We identified progression-related gene classifier that has strong predictive value for determining disease outcome in NMIBC. This gene classifier could assist in selecting NMIBC patients who might benefit from more aggressive therapeutic intervention or surveillance.

BACKGROUND:S100 calcium binding protein A8 (S100A8) has been implicated as a prognostic indicator in several types of cancer. However, previous studies are limited in their ability to predict the clinical behavior of the cancer. Here, we sought to identify a molecular signature based on S100A8 expression and to assess its usefulness as a prognostic indicator of disease progression in non-muscle invasive bladder cancer (NMIBC).METHODS:We used 103 primary NMIBC specimens for microarray gene expression profiling. The median follow-up period for all patients was 57.6 months (range: 3.2 to 137.0 months). Various statistical methods, including the leave-one-out cross validation method, were applied to identify a gene expression signature able to predict the likelihood of progression. The prognostic value of the gene expression signature was validated in an independent cohort (n = 302).RESULTS:Kaplan-Meier estimates revealed significant differences in disease progression associated with the expression signature of S100A8-correlated genes (log-rank test, P < 0.001). Multivariate Cox regression analysis revealed that the expression signature of S100A8-correlated genes was a strong predictor of disease progression (hazard ratio = 15.225, 95% confidence interval = 1.746 to 133.52, P = 0.014). We validated our results in an independent cohort and confirmed that this signature produced consistent prediction patterns. Finally, gene network analyses of the signature revealed that S100A8, IL1B, and S100A9 could be important mediators of the progression of NMIBC.CONCLUSIONS:The prognostic molecular signature defined by S100A8-correlated genes represents a promising diagnostic tool for the identification of NMIBC patients that have a high risk of progression to muscle invasive bladder cancer.

OBJECTIVES To investigate the association between expression levels of topoisomerase-II alpha (TOP-2 alpha) and prognosis in primary non-muscle-invasive bladder cancer (NMIBC). TOP-2 alpha, a marker of cell proliferation, has been assessed as a prognostic indicator in several types of cancer. However, currently available data on the role of TOP-2 alpha in prognosis are inconsistent.METHODS Top-2 alpha messenger ribonucleic acid (mRNA) levels were examined in 103 tumor specimens from patients with primary NMIBC by real-time polymerase chain reaction. Immunohistochemical staining was performed on 39 matched tumor samples. The median follow-up period for all patients was 51.8 months (range, 3.2-137).RESULTS The mRNA expression levels of TOP-2 alpha were significantly elevated in subjects with high-grade (P < .001) and high-stage (P = .041) tumors as compared with subjects with low-grade and low-stage tumors. Kaplan-Meier estimates revealed significant variation in tumor recurrence and progression depending on the level of TOP-2 alpha expression (log-rank test, P < .05).Multivariate Cox regression analysis revealed that the level of TOP-2 alpha expression is a strong predictor of recurrence (hazard ratio, 2.507; 95% confidence interval, 1.228-5.116; P = .012) and progression (hazard ratio, 4.192; 95% confidence interval, 1.002-17.536; P = .049) for NMIBC. The results of immunohistochemical staining generally correlated with mRNA expression levels.CONCLUSIONS Enhanced expression of TOP-2 alpha is positively associated with a high rate of recurrence and progression in NMIBC. Thus, TOP-2 alpha represents a promising marker of prognosis for NMIBC. UROLOGY 75: 1516.e9-1516.e13, 2010. (c) 2010 Elsevier Inc.

Purpose: DNA methylation is a key regulator of gene transcription and genomic stability, and alterations in DNA methylation are frequently detected in human tumors. Recent study has suggested that inactivation of runt-related transcription factor 3 (RUNX3), primarily epigenetic alterations in DNA methylation, is closely associated with bladder tumor stage, grade, and prognosis. The aim of this study was to evaluate the association between RUNX3 inactivation and renal cell carcinoma (RCC). Materials and Methods: RCC tissues (n=56) were obtained from patients who underwent radical nephrectomy. The methylation pattern of RUNX3 was determined by using methylation specific-polymerase chain reaction (MS-PCR) and direct DNA sequencing. Results: Methylation of the RUNX3 promoter was observed in 75.0% of the samples (42/56). The tumor stage and grade were significantly associated with the methylation status (p 0.05, respectively). Conclusions: This study demonstrated that promoter methylation of RUNX3 is frequently observed in RCC. In addition, RUNX3 methylation is closely associated with aggressive pathologic features. (Korean J Urol 2009;50:432-438) 󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏󰠏

A Case of Streptococcus salivarius Meningitis in a Patient with Cerebrospinal Fluid Rhinorrhea after Skull Base Fracture Kyeong Seob Shin, Dong Ik Shin, Woo Sub Shim, Byeong Cheol Rim, Il Hun Bae, Seung Young Lee, Dong Hee Ryu, Eun Jung Kim, Bo Ra Son Departments of Laboratory Medicine, Neurology, Otorhinolaryngology-Head and Neck Surgery, Neurosurgery, Radiology and Surgery, Chungbuk National University College of Medicine, Department of Pathology, Chungbuk National University Hospital, Cheongju, Korea

The expression of matrix metalloproteinase-9 (MMP-9) has been implicated in tumor invasion and metastasis. In this study, the factors and signaling pathways that are involved in the regulation of the MMP-9 expression were examined in urinary bladder cancer HT1376 cells. Tumor necrosis factor-alpha (TNF-alpha) stimulated the secretion of MMP-9 in HT1376 cells, as shown by zymography and immunoblot analysis. At the level of transcription, TNF-alpha also stimulated 5'-flanking promoter activity of MMP-9. Transcription factor NF-kappaB, AP-1 and Sp-1 binding sites were identified by a gel shift assay to be cis-elements for TNF-alpha activation of the MMP-9 promoter. TNF-alpha activates multiple signaling pathways in HT1376 cells, including the extracellular signal-regulated kinase (ERK1/2), p38 MAP kinase and JNK pathways. Chemical inhibitors, which specifically inhibit each of these TNF-alpha-activated pathways, were used to examine the signaling pathways involved in TNF-alpha-mediated MMP-9 expression. The ERK1/2 inhibitor, U0126 and the p38 MAP kinase inhibitor, SB203580, significantly down-regulated TNF-alpha-induced MMP-9 expression and promoter activity. The transactivation of TNF-alpha-stimulated NF-kappaB, AP-1 and Sp-1 were inhibited by U0126 and SB203580 treatment. In conclusion, the findings of the present study indicate that TNF-alpha induces MMP-9 expression in HT1376 cells by activating transcription factors, which are involved in the ERK1/2- and p38 MAP kinase-mediated control of MMP-9 regulation, namely, NF-kappaB, AP-1 and Sp-1.

Magnolol has been reported to play a role in antitumor activity. However, the relevant pathway integrating cell cycle regulation and signaling pathways involved in growth inhibition in cancer cells remains to be identified. In the present study, magnolol treatment of these cells resulted in significant dose-dependent growth inhibition together with apoptosis, G1- and G2/M-phase cell cycle arrest at a 60μM (IC50) dose in 5637 bladder cancer cells. In addition, magnolol treatment strongly induced p27KIP1 expression, and down-regulated expression of cyclin-dependent kinases (CDKs) and cyclins. Moreover, treatment with magnolol-induced phosphorylation of ERK, p38 MAP kinase, and JNK. Among the pathway inhibitors examined, only PD98059, an ERK-specific inhibitor, blocked magnolol-dependent p27KIP1 expression. Blockade of ERK function consistently reversed magnolol-mediated inhibition of cell proliferation and decreased G2/M cell cycle proteins, but not G1 cell cycle proteins. Furthermore, magnolol treatment increased both Ras and Raf activation. Transfection of cells with dominant negative Ras (RasN17) and Raf (RafS621A) mutant genes suppressed magnolol-induced ERK activity and p27KIP1 expression. Finally, the magnolol-induced reduction in cell proliferation and G2/M cell cycle proteins was also abolished in the presence of RasN17 and RafS621A mutant genes. These data demonstrate that the Ras/Raf/ERK pathway participates in p27KIP1 induction, leading to a decrease in the levels of cyclin B1/Cdc2 complexes and magnolol-dependent inhibition of cell growth. Overall, these novel findings concerning the molecular mechanisms of magnolol in 5637 bladder cancer cells provide a theoretical basis for therapeutic treatment of malignancies.

PURPOSE:DNA methylation is a key regulator of gene transcription and genomic stability, and alterations in DNA methylation patterns are frequently detected in human tumors. Previously we reported that inactivation of RUNX3 by primarily epigenetic alterations in DNA methylation is closely associated with bladder tumor development, recurrence and progression. In the current series we evaluated the association between RUNX3 inactivation and bladder tumors after a long-term followup study.MATERIALS AND METHODS:We used previously published data on the methylation patterns of RUNX3 in bladder tumor samples as well as 25 new data sets obtained by methylation specific polymerase chain reaction and direct DNA sequencing. Of the 149 patients examined 118 were followed periodically and included in the final analysis. Median followup was 49.8 months (range 1 to 146).RESULTS:RUNX3 promoter methylation was observed in 84 of the 118 tumor samples (71.2%) examined. RUNX3 methylation patterns correlated significantly with the development of invasive tumor, tumor progression, and overall and cancer specific survival (each p <0.05). Kaplan-Meyer curves showed identical results (p <0.05). Multivariate Cox regression models revealed that RUNX3 methylation status was a strong predictor of tumor progression and cancer specific survival.CONCLUSIONS:Results strongly suggest that inactivation of RUNX3 by the methylation of its promoter region is a significant risk factor for invasive bladder tumors, tumor progression and cancer specific survival. RUNX3 promoter methylation status could be a promising marker for assessing the prognosis of human bladder tumors.

You have accessJournal of UrologyDiscussed Poster, Monday, May 23, 2005, 8:00 am - 12:00 pm1 Apr 2005583: Transforming Growth Factor β1 and its Receptors are Related to the Development, Recurrence and Prgression of Human Bladder Cancers Wun-Jae Kim, ChangYi Quan, Pil-Du Jeoung, Eun-Jung Kim, Ji-Yeon Kim, Seok Hyung Kim, Suk-Chul Bae, and Joong-Kook Choi Wun-Jae KimWun-Jae Kim More articles by this author , ChangYi QuanChangYi Quan More articles by this author , Pil-Du JeoungPil-Du Jeoung More articles by this author , Eun-Jung KimEun-Jung Kim More articles by this author , Ji-Yeon KimJi-Yeon Kim More articles by this author , Seok Hyung KimSeok Hyung Kim More articles by this author , Suk-Chul BaeSuk-Chul Bae More articles by this author , and Joong-Kook ChoiJoong-Kook Choi More articles by this author View All Author Informationhttps://doi.org/10.1016/S0022-5347(18)34823-7AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail "583: Transforming Growth Factor β1 and its Receptors are Related to the Development, Recurrence and Prgression of Human Bladder Cancers." The Journal of Urology, 173(4S), p. 159 © 2016 by American Urological AssociationFiguresReferencesRelatedDetails Volume 173Issue 4SApril 2005Page: 159 Advertisement Copyright & Permissions© 2016 by American Urological AssociationMetricsAuthor Information Wun-Jae Kim More articles by this author ChangYi Quan More articles by this author Pil-Du Jeoung More articles by this author Eun-Jung Kim More articles by this author Ji-Yeon Kim More articles by this author Seok Hyung Kim More articles by this author Suk-Chul Bae More articles by this author Joong-Kook Choi More articles by this author Expand All Advertisement PDF DownloadLoading ...

RUNX3 is inactivated at high frequency in many tumors. However, in most cases, inactivation is caused by silencing of the gene due to promoter hypermethylation. Because epigenetic silencing is known to affect many major tumor suppressor genes in cancer cells, it is not clear whether RUNX3 is primarily responsible for the induction of carcinogenesis in these cases, except for the gastric cancer cases that we reported previously. We investigated genetic and epigenetic alterations of RUNX3 in 124 bladder tumor cases and seven bladder tumor-derived cell lines. Here we show that RUNX3 is inactivated by aberrant DNA methylation in 73% (90 of 124) of primary bladder tumor specimens and 86% (six of seven) of bladder tumor cell lines. In contrast, the promoter regions of 20 normal bladder mucosae were unmethylated. Importantly, one patient bore missense mutations, each of which resulted in amino acid substitutions in the highly conserved Runt domain. The mutations abolished the DNA-binding ability of RUNX3. A second patient had a single nucleotide deletion within the Runt domain coding region that resulted in truncation of the protein. RUNX3 methylation was a significant risk factor for bladder tumor development, superficial bladder tumor recurrence, and subsequent tumor progression. These results strongly suggest that inactivation of RUNX3 may contribute to bladder tumor development and that promoter methylation and silencing of RUNX3 could be useful prognostic markers for both bladder tumor recurrence and progression.