Yutaka Horiguchi

Prognostic value of neutrophil-to-lymphocyte ratio and establishment of novel preoperative risk stratification model in bladder cancer patients treated with radical cystectomy.

OBJECTIVEnPreoperative prognostic factors in bladder cancer patients have not been fully established. This study was undertaken to investigate preoperative prognostic factors, including neutrophil-to-lymphocyte ratio (NLR), and to develop a novel prognostic factors-based risk stratification model for disease-specific survival (DSS) in bladder cancer patients treated with radical cystectomy (RC).nnnMETHODSnWe performed a retrospective analysis of 189 consecutive bladder cancer patients treated with RC at our institution. Prognostic value of the preoperative clinical and laboratory parameters were evaluated by univariate and multivariate Cox proportional hazard model analyses, and patients were stratified according to relative risks (RRs) for DSS.nnnRESULTSnOne-, 3-, and 5-year DSS rates were 86.8%, 70.8%, and 61.7%, respectively. In univariate analysis, tumor size, clinical T stage, hydronephrosis, concomitance of carcinoma in situ, and some laboratory findings (hemoglobin [Hb] level, platelet count, C-reactive protein, neutrophil count, lymphocyte count, and NLR) were significantly associated with poor prognosis. In multivariate analysis, tumor size, hydronephrosis, Hb level, and NLR were independent factors for predicting poor prognosis. Patients were stratified into 3 risk groups: low (RR = 1.000-3.717), intermediate (RR = 4.149-9.315), and high (RR = 10.397-38.646). The differences among the groups were significant.nnnCONCLUSIONSnNLR was an independent prognostic factor, as were tumor size, hydronephrosis, and Hb levels, and the combination of these factors can stratify DSS risks in bladder cancer patients treated with RC. This information may be useful for identifying patients who might be candidates for clinical trials of multimodal treatment strategies, including innovative neoadjuvant treatments.

Comparison of CT Urography and Excretory Urography in the Detection and Localization of Urothelial Carcinoma of the Upper Urinary Tract

OBJECTIVEnThe purpose of this study was to compare the accuracy of CT urography and excretory urography for the detection and localization of upper urinary tract urothelial carcinoma.nnnMATERIALS AND METHODSnOf 128 patients at high risk for upper tract urothelial carcinoma who were examined with both CT urography and excretory urography between 2002 and 2007, 24 were undiagnosed and excluded. CT urography and excretory urography results of the remaining 104 patients and 552 urinary tract segments were compared with histopathologic examination or follow-up imaging at 1 year. Two readers independently scored the confidence levels for the presence or absence of upper urinary tract urothelial carcinoma in each of six upper urinary tract segments on both CT urography and excretory urography; differences were resolved by consensus.nnnRESULTSnUpper urinary tract urothelial carcinoma was diagnosed in 77 (14%) segments of 46 (44%) patients. Per-patient sensitivity, specificity, overall accuracy, and area under the receiver operating characteristic curves for detecting carcinomas with CT urography (93.5% [43/46], 94.8% [55/58], 94.2% [98/104], and 0.963, respectively) were significantly greater than those for excretory urography (80.4% [37/46], 81.0% [47/58], 80.8% [84/104], and 0.831, respectively) (p = 0.041, p = 0.027, p = 0.001, and p < 0.001, respectively). Per-segment sensitivity and overall accuracy for the localization of upper urinary tract urothelial carcinoma were significantly greater with CT urography (87.0% [67/77] and 97.8% [540/552]) than with excretory urography (41.6% [32/77] and 91.5% [505/552]) (p < 0.0001).nnnCONCLUSIONnCT urography was more accurate than excretory urography in the detection and localization of upper urinary tract urothelial carcinoma and should be considered as the initial examination for the evaluation of patients at high risk for upper urinary tract urothelial carcinoma.

Detection of Bladder Tumors with Dynamic Contrast-Enhanced MDCT

OBJECTIVEnIn a small pilot study, we assessed whether early-phase dynamic contrast-enhanced MDCT can be used to detect bladder tumors and whether thin reconstruction improves the detection rate.nnnSUBJECTS AND METHODSnThirty-six patients (30 with 59 cystoscopy-proven bladder cancers and six with normal bladders) underwent dynamic contrast-enhanced MDCT of the pelvis and abdomen. Images were obtained from the symphysis pubis to the diaphragm 70 seconds after injection of 100 mL of contrast medium. McNemar test was used to compare sensitivity per patient, segment, and tumor and specificity per patient and segment for each of three reconstruction methods: 5-mm sections with no overlap (i.e., 5-mm axial images), 2.5-mm sections with 1.25-mm overlap (i.e., thin-section axial images), and 2.5-mm sections with 1.25-mm overlap and multiplanar reformation (MPR) (i.e., thin-section axial images with MPR).nnnRESULTSnMDCT with a combination of thin, overlapped sections and MPR depicted all but one of 47 bladder tumors larger than 5 mm but only five of 12 tumors 5 mm or smaller. There were no false-positive findings. Per-tumor sensitivity was significantly better with thin-section images with MPR (90%) and thin-section images alone (86%) than with 5-mm axial images (80%) (p < 0.05). Per-segment sensitivity was significantly better with thin-section images with MPR (95%) and thin-section axial images alone (87%) than with 5-mm axial images (79%) (p < 0.05). Per-patient sensitivity and per-patient and per-segment specificity did not differ with the three methods.nnnCONCLUSIONnDynamic contrast-enhanced MDCT of the pelvis shows promise for the detection of bladder tumors. Use of thin-section images with MPR and thin-section axial images alone had a significantly better rate of detection of bladder tumors than use of 5-mm axial images.

Effects of a KiSS-1 peptide, a metastasis suppressor gene, on the invasive ability of renal cell carcinoma cells through a modulation of a matrix metalloproteinase 2 expression

Although effects of a metastasis suppressor gene, KiSS-1, have been postulated to be mediated by its receptor, hOT7T175, the mechanism of such effects remains unknown. This study was designed to evaluate the mechanism of how KiSS-1 works and to assess effects of a synthesized truncated KiSS-1 protein on the invasive ability of renal cell carcinoma (RCC) cells. Four RCC cell lines, Caki-1, KU19-20, RSP and RSM, were investigated to determine mRNA expressions of KiSS-1, its receptor, hOT7T175, matrix metalloproteinases (MMPs) and MMP inhibitors. While all cell lines demonstrated hOT7T175 mRNA expressions, only Caki-1 had KiSS-1 transcripts. A synthesized truncated KiSS-1 peptide, metastin (45-54), produced a marked suppression of the invasive ability in KU19-20 cells, which were deficient for KiSS-1 transcripts, but not in Caki-1 cells. Metastin (45-54) also increased the ability of KU19-20 cells to attach to collagen 4. Both MMP-2 mRNA levels and protein production were significantly decreased only in KU19-20 cells by metastin (45-54). In conclusion, metastin (45-54) may have potential therapeutic use by suppressing the motility and invasive ability of RCC cells which possess hOT7T175 with either a negative expression or very low expression level of KiSS-1 through, at least in part, the down-regulation of MMP-2.

Hiding in plain view: Genetic profiling reveals decades old cross contamination of bladder cancer cell line ku7 with hela

PURPOSEnKU7 is a popular urothelial carcinoma cell line that was isolated from the bladder of a patient at Keio University in 1980. It has subsequently been widely used in laboratories around the world. We describe how routine cell line authentication revealed that KU7 was cross contaminated almost 30 years ago with HeLa, a cervical carcinoma cell line.nnnMATERIALS AND METHODSnPresumed KU7 clones dating from 1984 to 1999 were provided by M.D. Anderson Cancer Center, Vancouver Prostate Centre, Kyoto University, Tokyo Medical University and Keio University. HeLa was obtained from ATCC. Genomic DNA was isolated and short tandem repeat analysis was performed at the M.D. Anderson Cancer Center Characterized Cell Line Core Facility, Johns Hopkins University Fragment Analysis Facility and RIKEN BioResource Center, Ibaraki, Japan. Comparative genomic hybridization was performed on a platform (Agilent Technologies, Santa Clara, California) at Vancouver Prostate Centre.nnnRESULTSnThe short tandem repeat profile of all KU7 clones was an exact match with that of HeLa. Comparative genomic hybridization of all samples revealed an abundance of shared chromosomal aberrations. Slight differences in some genomic areas were explained by genomic drift in different KU7 clones separated by many years.nnnCONCLUSIONSnOur analysis identified that cross contamination of KU7 with HeLa occurred before 1984 at the source institution. All KU7 clones in the urological literature should be considered HeLa and experimental results should be viewed in this light. Our results emphasize the need to authenticate cell lines in oncological research.

Angiotensin II Type 1 Receptor Antagonist Enhances Cis-dichlorodiammineplatinum-Induced Cytotoxicity in Mouse Xenograft Model of Bladder Cancer

OBJECTIVESnTo examine whether candesartan enhances the cytotoxicity of cis-dichlorodiammineplatinum (CDDP) in mice with bladder cancer as a method to enhance the therapeutic effects of CDDP. CDDP is an antitumor agent conventionally used against bladder cancer; however, its therapeutic efficacy appears to not be fully satisfactory. Recent studies have shown the antitumor activity of the angiotensin II type 1 receptor antagonist candesartan.nnnMETHODSnA xenograft model was prepared in nude mice using human bladder cancer cells (KU-19-19). Candesartan (1 mg/kg/d) was administered daily by oral gavage from the day of implantation plus 28 days, and CDDP (1 mg/kg/d) was administered intraperitoneally from days 5 to 9. The microvessel density, vascular endothelial growth factor expression, and apoptosis were investigated immunohistochemically.nnnRESULTSnCandesartan, CDDP, and candesartan-CDDP suppressed tumor growth to 41.9%, 33.8%, and 13.2%, respectively, of the tumor volume in the control group, showing that combined treatment significantly inhibited tumor growth compared with each single agent alone. The microvessel density was significantly decreased in the candesartan and candesartan-CDDP groups compared with the control group. Vascular endothelial growth factor expression was significantly decreased in the candesartan, CDDP, and candesartan-CDDP groups compared with the control group. The apoptotic index was significantly increased in the CDDP and candesartan-CDDP groups compared with the control and candesartan groups.nnnCONCLUSIONSnIt is quite likely that candesartan and CDDP suppressed tumor growth by inhibiting angiogenesis and inducing apoptosis, respectively. Furthermore, combined treatment with candesartan enhanced CDDP-induced cytotoxicity by further suppressing angiogenesis. These results suggest that candesartan could be a candidate for innovational therapy of bladder cancer.

Enhancement of radiosensitivity by a unique novel NF-κB inhibitor, DHMEQ, in prostate cancer

Background:Inducible activation of nuclear factor (NF)-κB is one of the principal mechanisms through which resistant prostate cancer cells are protected from radiotherapy. We hypothesised that inactivation of inducible NF-κB with a novel NF-κB inhibitor, DHMEQ, would increase the therapeutic effects of radiotherapy.Methods:PC-3 and LNCaP cells were exposed to irradiation and/or DHMEQ. Cell viability, cell cycle analysis, western blotting assay, and NF-κB activity were measured. The antitumour effect of irradiation combined with DHMEQ in vivo was also assessed.Results:The combination of DHMEQ with irradiation resulted in cell growth inhibition and G2/M arrest relative to treatment with irradiation alone. Inducible NF-κB activity by irradiation was inhibited by DHMEQ treatment. The expression of p53 and p21 in LNCaP, and of 14-3-3σ in PC-3 cells, was increased in the combination treatment. In the in vivo study, 64 days after the start of treatment, tumour size was 85.1%, 77.1%, and 64.7% smaller in the combination treatment group than that of the untreated control, DHMEQ-treated alone, and irradiation alone groups, respectively.Conclusion:Blockade of NF-κB activity induced by radiation with DHMEQ could overcome radio-resistant responses and may become a new therapeutic modality for treating prostate cancer.

Production of erythropoietin and multiple cytokines by metanephric adenoma results in erythrocytosis

This is the first report of direct evidence that metanephric adenoma cells produce erythropoietin and other types of cytokines, which may be the cause of the high incidence of erythrocytosis in patients with this tumor. The purpose of the study was to establish a metanephric adenoma cell line in vitro from nephrectomized tumor tissue in order to investigate the ability of metanephric adenoma cells to produce erythropoietin and other types of cytokines. The tumor tissue was obtained from a 16‐year‐old boy who had developed metanephric adenoma with erythrocytosis and was served for cell culture. Significantly high concentrations of erythropoietin, granulocyte–macrophage colony‐stimulating factor (GM‐CSF), granulocyte–colony‐stimulating factor (G‐CSF), interleukin‐6 (IL‐6), and IL‐8 were detected in the cell culture supernatant. Southern hybridization showed specific positive signals for GM‐CSF, G‐CSF, IL‐6, IL‐8 and erythropoietin. The number of chromosomes was 46‐XY without any structural abnormalities in cytogenetic analysis of the cultured cells.

The Novel NF-κB Activation Inhibitor Dehydroxymethyl-Epoxyquinomicin Suppresses Anti-Thy1.1-Induced Glomerulonephritis in Rats

Abstract Background: NF-κB participates in the transcriptional regulation of numerous genes, and many studies have confirmed the activation of NF-κB in inflammatory renal diseases. Therefore, NF-κB is a promising target for the treatment of these diseases. We tested the effects of dehydroxymethyl-epoxyquinomicin (DHMEQ), a novel NF-κB activation inhibitor, on anti-thy1.1 antibody-induced glomerulonephritis (Thy1.1 GN). Methods: Thy1.1 GN was induced in Sprague-Dawley rats (6/group) by intravenous injection of anti-thy 1.1 antibody. The effects of DHMEQ (8 mg/kg/day) on the glomerular disease were evaluated using periodic acid-Schiff and Masson trichrome stains, immunohistochemistry for proliferating cell nuclear antigen, fibronectin and CD45 (leukocyte common antigen) and TUNEL staining. NF-κB activation was analyzed by a fluorescent electrophoretic mobility shift assay. Results: On day 7, DHMEQ treatment resulted in marked inhibition of NF-κB, decreased proteinuria (223.2 ± 42.3 vs. 434.8 ± 16.5 mg/kg/day, p < 0.05), preserved creatinine clearance (1.93 ± 0.38 vs. 1.07 ± 0.29 l/day, p < 0.01), decreased glomerular cell proliferation (15.8 ± 1.2 vs. 31.2 ± 0.8 nuclei/glomerular cross-section) and mesangial matrix deposition, and an increase in glomerular and tubular apoptosis without inducing any obvious adverse effects. Conclusion: DHMEQ inhibited NF-κB and thereby suppressed the inflammatory renal responses in rats with Thy1.1 GN.

Risk stratification of survival by lymphovascular invasion, pathological stage, and surgical margin in patients with bladder cancer treated with radical cystectomy

BackgroundThe aim of this study was to investigate prognostic factors and develop a prognostic factor-based risk stratification model for disease-specific survival (DSS) in a radical cystectomy (RC) series.MethodsThe patient cohort comprised 194 consecutive patients with bladder cancer treated with RC. Univariate and multivariate Cox proportional hazard model analyses were performed to identify significant prognostic factors for DSS. A risk stratification model was developed based on the relative risks (RRs) of DSS.ResultsMedian follow-up period was 26.8xa0months. The 1-, 3-, and 5-year DSS were 88.0, 74.0, and 64.9%, respectively. In the univariate analysis, pathological T (pT) (≥pT2), lymphovascular invasion (LVI), non-urothelial carcinoma component, surgical margin (SM), and lymph node metastases (pN) were significantly associated with poor prognosis. In the multivariate analysis, pT (≥pT2), LVI, and SM were independent factors for predicting poor prognosis. Based on these results, patients were stratified into three risk groups: low (RRxa0=xa01.00–3.626), intermediate (5.860–9.826), and high (21.24). The 1-, 3-, and 5-year survival rates were 96.9, 85.1, and 85.1% in the low-risk group, 83.0, 63.4, and 43.8% in the intermediate group, and 51.0, 19.4, and 19.4% in the high-risk group, respectively. The differences among these groups were significant.ConclusionsIn our RC series, pT (≥pT2), LVI, and SM were independent prognostic factors. This information may be useful to identify patients with poor prognosis, who might be good candidates for innovative treatment.