Muscle-invasive Cancer Research Articles

Abstract 3832: Cancer tissue-derived spheroid, CTOS, for evaluation of drug response from individual patient tumor samples.

Abstract Primary culture of cancer cells can be an excellent platform for investigating the biology of cancer and predicting chemosensitivity for individual patients, yet has been hampered by technical difficulties. We established a method of primary culture for human colorectal cancer: cancer tissue-originated spheroid (CTOS) method. The principle of the method is to maintain cell-cell contact throughout preparation and culture; never dissociating the samples into single cells. The clusters of cancer cells turn out to spheroids, CTOSs, within a short period. CTOSs of colorectal cancer consisted of highly purified cancer cells, and they were prepared with high efficiency. We applied the CTOS method to non-small cell lung cancer and urothelial cancer. For non-small cell lung cancer, we used 125 surgical specimens and 18 pleural effusions. The success rate of CTOS preparation from surgical specimens was 80.0%. CTOSs were subjected to sensitivity assay and pathway activation assay for the EGFR tyrosine kinase inhibitor (TKI) erlotinib. The sensitivity differed among individual patient CTOSs. The EGFR mutation status and expression levels corresponded with erlotinib sensitivity, confirming previous clinical findings. Next, we examined urothelial cancer. The success rate of CTOS preparation from the non muscle-invasive urothelial cancer was 90.7%, whereas that from the muscle-invasive urothelial cancer was 68.2%. Total development rate was 84.2%. Using these CTOSs, we investigated the effects of growth factors on the growth of CTOS in vitro. Among the growth factors examined, including EGF, IGF-1, basic FGF, and activin A, we found that heregulin (HRG)/neuregulin1, a ligand of HER3, potently induced CTOS growth in both lung and urothelial cancers. HRG promoted the growth of primary CTOSs in 10 out of 13 (71.4%) cases of lung cancer, and in 2 out of 4 (50%) cases of urothelial cancer. Along with the growth, HRG most potently stimulated the phosphorylation of AKT. In lung cancer CTOSs, a neutralizing antibody against HER3 inhibited the growth and the phosphorylation of HER3 and AKT. In all three tumor types examined, CTOSs formed xenograft tumors in immunodeficient mice, and the tumors retained the features of the parental tumors. As CTOS can be cryopreserved, we are accumulating stocks of CTOSs derived from patient samples as well as the CTOSs from xenograft tumors. CTOS experiments can be shuttled between in vitro and in vivo. Thus, the CTOS method enables us to obtain primary colorectal, lung, and urothelial tumor cells of high viability and purity. CTOS could be a new platform for studying lung cancer biology, and be useful to select patients for molecular targeting drugs. Citation Format: Masahiro Inoue, Hiroko Endo, Masayuki Ohue, Kazuo Nishimura, Fumio Imamura, Jiro Okami, Hiroaki Okuyama. Cancer tissue-derived spheroid, CTOS, for evaluation of drug response from individual patient tumor samples. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3832. doi:10.1158/1538-7445.AM2013-3832

High level of mature tumor-infiltrating dendritic cells predicts progression to muscle invasion in bladder cancer

A prognostic value for tumor-associated macrophages (TAMs) and tumor-infiltrating dendritic cells (TIDCs) has been reported in many human cancers. The objective of this study is to determine the prognostic value of CD83(+) mature TIDCs and CD68(+) TAMs in non-muscle-invasive bladder cancer at first diagnosis. Immunohistochemistry staining was performed with anti-CD68 and anti-CD83 monoclonal antibodies on tissue sections from 93 formalin-fixed, paraffin-embedded tissue blocks from pTa and pT1 bladder tumors. A scoring index based on the average density of observed positive cells in the papillary axis, the stroma, lymphoid aggregates, and into tumor foci was calculated for each patient. Comparison of baseline characteristics with marker levels was done using Pearson χ(2) or Fisher exact test. Kaplan-Meier analyses and Cox regression models were fitted to evaluate the prognostic value of TIDCs and TAMs. The absence of both CD68(+) TAMs and CD83(+) TIDCs was associated with tumor recurrence. The presence of TIDCs was associated with a significant risk of progression to muscle-invasive cancer (hazard ratio, 8.253; P = .0179). Patients were risk stratified using age and TIDC score. Patients 70 years or older with a high score of TIDCs had a 56% progression-free survival after 6 years compared with 94% for patients younger than 70 years with a low score of TIDCs. None of 20 patients with a low score of TAMs progressed. These data indicate that the presence of mature TIDCs and possibly TAMs may help risk-stratify patients at the time of first diagnosis of non-muscle-invasive bladder cancer and may be useful in tailoring follow-up and treatment strategies.

Radiologic response (RR) by MRI in patients (pts) with muscle-invasive urothelial cancer (MIUC) undergoing neoadjuvant chemotherapy.

283 Background: Standard response criteria do not always provide a meaningful assessment of treatment benefit in pts with MIUC. We examined the utility of contrast-enhanced imaging in predicting disease free survival (DFS) in MIUC pts, and its correlation with pathological downstaging on radical cystectomy (RC) specimen. Methods: Baseline and post chemotherapy imaging, primarily gadolinium-enhanced pelvic MRI, from pts enrolled on a phase II prospective study of neoadjuvant cisplatin-based chemotherapy were reviewed by 2 oncoradiologists (NR, KK). DFS was defined as time from RC to evidence of disease progression or death. Qualitative (QL) RR was defined by at least moderate decrease in the primary tumor and lymph nodes after chemotherapy, with visually decreased primary tumor bulk by at least half, normalization of node size, and delayed enhancement of residual tumor and nodes. Quantitative (QN) RR was assessed according to anatomic TNM downstaging per AJCC staging criteria 7.0. Fisher’s exact test was used to compare RR with pathological dowstaging. DFS distributions within RR subgroups were estimated with the Kaplan-Meier method and compared using the logrank test. Results: All 39 pts from the original study were included in this analysis. At baseline, 42% of pts had cT2 disease, 42% had cT3, 16% had cT4, and 45% had N1 disease. 91% of pts had bladder primary, and 84% of pts were assessed by MRI. Results showed QL RR in 64% (90% CI 50-77) of pts, QN RR in 61% (90% CI 47-75), and ≤pT1N0M0 in 49% (90% CI 35-63). While there was no association between pathological downstaging and QN RR, there was a modest association with QL RR (p=0.096). 60% of pts with QL RR achieved ≤pT1N0M0 and 71% of pts without QL RR were >pT2. QL RR was also a strong predictor of DFS with an 18-month DFS advantage of 83% (95% CI 56-94) among responders vs. 32% (95% CI 6-63) in non-responders (p=0.036). Conclusions: QL RR post neoadjuvant therapy is modestly associated with pathological downstaging in pts with MIUC and strongly predictive of DFS. Results suggest post-chemotherapy, pre-operative imaging may be a valuable prognostic tool. Confirmation in other UC populations and treatment settings should be explored.

Involvement of Heregulin/HER3 in the Primary Culture of Human Urothelial Cancer

We previously established a novel method of human colorectal cancer primary culture. This method, termed the cancer tissue originated spheroid method, involves the preparation of multicellular spheroids of primary cancer cells that are cultured so that cell-cell contact is maintained. We applied this method to human urothelial cancer. Cancer tissue originated spheroids were prepared from xenografts or primary human bladder urothelial cancer tumors following the same protocol used for human colorectal cancer. Cancer tissue originated spheroids were characterized using immunohistochemistry, Western blot and polymerase chain reaction. We established a xenograft from a primary bladder urothelial cancer, and isolated and cultured cancer tissue originated spheroids from the xenograft tumor. Cancer tissue originated spheroids retained the characteristics of the original tumor and those of the xenograft. Heregulin promoted cancer tissue originated spheroid growth, and inhibitors of PI3K and mTOR inhibited heregulin induced growth, as did lapatinib but not erlotinib. We also prepared cancer tissue originated spheroids from primary bladder urothelial cancer. The success rate of establishing primary cancer tissue originated spheroids from nonmuscle invasive urothelial cancer was 90.7% and that from muscle invasive cancer was 68.2%. The overall success rate was 84.2%. Heregulin promoted the growth of primary cancer tissue originated spheroids from 4 of 7 patients. We report a method of establishing primary cultures of human urothelial cancer cells. Growth stimulation by heregulin in cancer tissue originated spheroids from xenografts and primary tumors suggests the possibility of molecular targeting therapy against HER3 signaling for human urothelial cancer. The cancer tissue originated spheroid method might be useful for selecting patients for molecular targeting drugs such as lapatinib.

Urinary Bladder Sinuses

We described urinary bladder sinuses (UBS) in the urinary bladder, characterized by segmental mucosal with muscularis mucosa invaginations into the submucosa (superficial UBS) and muscularis propria (deep UBS). Radical cystectomy specimens and transurethral resections of the bladder were reviewed. Superficial UBS (extending up to the submucosa) were distinguished from cystitis cystica, and deep UBS differed from embryonic remnants, intramural ureters, and diverticulum by their cleft-like structures lined by the urothelium. In transurethral resections of the bladder specimens and bladder biopsy specimens, superficial UBS were identified in 13/20 cases. In radical cystectomy specimens, superficial UBS were identified in 15/50 cases, whereas deep UBS (all with associated superficial UBS) were seen in 23/50 additional cases. Deep UBS limited to the inner quarter of the muscularis propria and only reached the level of the inner third outer/middle third of the muscularis propria in 5 cases. Intraepithelial neoplasia involved the mucosa of UBS in 10 cases; this may pose diagnostic problems with invasive carcinoma. Furthermore, UBS were often associated with mucosal redundancy and hypertrophy of the muscularis propria, therefore mimicking muscle invasive cancer on pelvic examination and imaging techniques. Recognition of UBS is important to pathologically and clinically avoid over-staging of bladder malignancies.

A phase 2 clinical trial of sequential neoadjuvant chemotherapy with ifosfamide, doxorubicin, and gemcitabine followed by cisplatin, gemcitabine, and ifosfamide in locally advanced urothelial cancer

Neoadjuvant chemotherapy improves the survival of patients with high-risk urothelial cancer. However, the lack of curative alternatives to cisplatin-based chemotherapy is limiting for patients with neuropathy or hearing loss. Sequential chemotherapy also has not been well studied in the neoadjuvant setting. The authors explored sequential neoadjuvant ifosfamide-based chemotherapy in a patient cohort at high risk of noncurative cystectomy. Patients with muscle-invasive cancer and lymphovascular invasion, hydronephrosis, clinical T3b and T4a (cT3b-4a) disease (defined as a 3-dimensional mass on examination under anesthetic or invasion into local organs), micropapillary tumors, or upper tract disease received 3 cycles of combined ifosfamide, doxorubicin, and gemcitabine followed by 4 cycles of combined cisplatin, gemcitabine, and ifosfamide. The primary endpoint was downstaging to pT1N0M0 disease or lower. At a median follow-up of 85.3 months, the 5-year overall survival (OS) and disease-specific survival (DSS) rates for all 65 patients were 63% and 68%, respectively (95% confidence interval: 5-year OS rate, 0.52%-0.76%; 5-year DSS rate, 0.58%-0.81%). Pathologic downstaging to pT1N0 disease or lower occurred in 50% of patients who underwent cystectomy and in 60% of patients who underwent nephroureterectomy and was correlated with the 5-year OS rate (pT1N0 disease or lower, 87%; pT2-pT3aN0 disease, 67%; and pT3b disease or higher or lymph node-negative disease, 27%; P ≤ .001 for pT1 or lower vs pT2 or higher). Variant histology was associated with an inferior 5-year DSS rate (50% vs 83% in pure transitional cell carcinoma; P = .02). The most frequent grade 3 toxicities were infection (38%), febrile neutropenia (22%), and mucositis (18%). There were 3 grade 4 toxicities (myocardial infarction, thrombocytopenia, and vomiting) and 1 grade 5 toxicity in a patient who refused antibiotics for pneumonia. Sequential therapy was active and maintained the historic expectation of achieving a cure. The current results strongly reinforced previous experience suggesting that pathologic downstaging to pT1N0 disease or less is a useful surrogate for eventual cure in patients with urothelial cancer.

Reduced CD151 expression is related to advanced tumour stage in urothelial bladder cancer

CD151 belongs to the group of tetraspanins and is aberrantly expressed in different tumours and differential expression has been associated with prognosis. The aim of this study was to clarify the relationship of CD151 expression with tumour phenotype and clinical outcome in bladder cancer. A bladder cancer tissue microarray containing samples from 686 urothelial bladder cancers was analysed by immunohistochemistry. Membranous CD151 immunostaining was recorded in 409 (66.0%) of 620 analysable cases. High CD151 expression was seen in normal urothelium and in most non-invasive tumours. Low CD151 expression levels were associated with a more unfavourable tumour phenotype. CD151 staining was seen in 71.5% of 284 pTa, 62.1% of 145 pT1 and 60.4% of 187 pT2-4 cancers (p = 0.0033). CD151 staining was detectable in 77.3% of 75 grade 1, 71.1% of 273 grade 2 and 57.7% of 272 grade 3 cancers (p < 0.0001). CD151 expression status was not associated with overall or tumour-specific survival in muscle-invasive cancers (pT2-4), tumour progression in pT1 and recurrences in pTa tumours. On the basis of our data we conclude that loss of CD151 may contribute to bladder cancer progression through attenuation of cell adhesion. In clinically defined subgroups CD151 expression does not provide additional prognostic information.

Stat3 Activation in Urothelial Stem Cells Leads to Direct Progression to Invasive Bladder Cancer

Two subtypes of human bladder cancer, noninvasive papillary and muscle-invasive cancer, develop through independent pathologic and molecular pathways. Human invasive bladder cancer frequently develops without prior clinical evidence of a noninvasive tumor stage. However, an animal model that recapitulates this unique clinical progression of invasive bladder cancer has not yet been developed. In this study, we created a novel transgenic mouse model of invasive bladder cancer by targeting an active dimerized form of Stat3 to the basal cells of bladder epithelium. When exposed to the carcinogen nitrosamine, Stat3-transgenic mice developed invasive cancer directly from carcinoma in situ (CIS), bypassing the noninvasive papillary tumor stage. Remarkably, invasive bladder cancer driven by active Stat3 was predominantly composed of stem cells, which were characterized by cytokeratin 14 (CK14) staining and enhanced tumor sphere-forming ability. Active Stat3 was also shown to localize to the nucleus of human invasive bladder cancers that were primarily composed of CK14+ stem cells. Together, our findings show that Stat3-induced stem cell expansion plays a critical role in the unique clinical progression of invasive bladder cancer through the CIS pathway.

Role of magnetic resonance imaging in bladder cancer: current status and emerging techniques

To evaluate the current status of magnetic resonance imaging (MR) as a staging tool for bladder cancer. To investigate the role of MR in assessing chemotherapeutic response in bladder cancer patients. A Pubmed/MEDLINE search was conducted to identify original articles, review articles, and editorials regarding the use of MR in bladder cancer. Contrast-enhanced MR and diffusion weighted MR (DW-MRI) can likely distinguish between non-muscle invasive bladder cancer and muscle invasive cancer with >80% accuracy. Some advantages of DW-MRI are the differentiation of benign versus malignant tissue involvement without the need for intravenous contrast, and the possibility of obtaining information on histologic grade and T stage. Traditional MR sequence have low sensitivity for identifying small lymph node metastases but MR lymphography (MRL) using ultra-small paramagnetic iron oxide (USPIO) may enhance their detectin. There may be a role for DW-MRI in the evaluation of chemotherapeutic response in bladder cancer patients. To date, sample sizes and study designs are insufficient to clearly establish the role of MR in bladder cancer management, and to this end, well designed prospective trials are needed.

890 INCREASED EXPRESSION OF SERUM UROPLAKIN III IS ASSOCIATED WITH THE DETECTION AND PATHOLOGICAL FEATURES OF AGGRESSIVE BLADDER CANCER

You have accessJournal of UrologyBladder Cancer: Basic Research II1 Apr 2012890 INCREASED EXPRESSION OF SERUM UROPLAKIN III IS ASSOCIATED WITH THE DETECTION AND PATHOLOGICAL FEATURES OF AGGRESSIVE BLADDER CANCER Kazumasa Matsumoto, Toshihide Matsumoto, Takefumi Satoh, Masatsugu Iwamura, Yuichi Sato, and Shiro Baba Kazumasa MatsumotoKazumasa Matsumoto Sagamihara, Japan More articles by this author , Toshihide MatsumotoToshihide Matsumoto Sagamihara, Japan More articles by this author , Takefumi SatohTakefumi Satoh Sagamihara, Japan More articles by this author , Masatsugu IwamuraMasatsugu Iwamura Sagamihara, Japan More articles by this author , Yuichi SatoYuichi Sato Sagamihara, Japan More articles by this author , and Shiro BabaShiro Baba Sagamihara, Japan More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.985AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The asymmetric unit membrane is a unique plasma membrane produced by differentiated urothelial cells, which mainly comprises the four transmembrane proteins uroplakin Ia, Ib, II and III. Owing to their urothelium-specific expression, the uroplakins have been widely investigated as potential markers for the immunohistochemical staining of primary lesions, the identification of primary cancers of unknown origin in patients with metastases and gene therapy for bladder cancer; however, the role of uroplakin proteins in the bladder remains unknown. This study detected preoperative levels of uroplakin III in the serum of patients with urothelial carcinoma of the bladder, and examined whether the expression of this protein could predict disease progression and survival. METHODS Uroplakin III expression was measured in the serum of 52 patients with bladder urothelial carcinoma, 28 normal controls and 13 patients with other diseases including renal cell carcinomas, adrenal lesions and acute pyelonephritis. The expression of uroplakin III was correlated with the pathological features and clinical outcomes. Anti-uroplakin III antibody was detected in the serum using the automated dot blot system and a micro-dot blot array with a 256 solid-pin system. RESULTS Uroplakin III expression was significantly increased in urothelial carcinoma patients compared with normal controls and patients with other diseases (p=0.0004); there were no significant differences between the latter two groups. Increased uroplakin III expression was associated with high-grade and muscle-invasive cancers (p=0.01 and 0.0004, respectively). After adjusting for the effects of standard pathological features, only muscle-invasive cancer was found to be associated with bladder cancer progression (p=0.003) and mortality (p=0.01). CONCLUSIONS Serum uroplakin III expression was significantly higher in patients with bladder cancer than in normal controls and patients with other diseases. Incremental uroplakin III expression was also associated with established features of biologically aggressive bladder cancer such as pathological stage and grade. Although uroplakin III has limited prognostic value, its expression in serum could be a valuable marker for the detection of bladder cancer. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e362 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Kazumasa Matsumoto Sagamihara, Japan More articles by this author Toshihide Matsumoto Sagamihara, Japan More articles by this author Takefumi Satoh Sagamihara, Japan More articles by this author Masatsugu Iwamura Sagamihara, Japan More articles by this author Yuichi Sato Sagamihara, Japan More articles by this author Shiro Baba Sagamihara, Japan More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

1781 WHAT IS THE RISK OF HIGH RISK NON-MUSCLE INVASIVE BLADDER CANCER? AN ENGLISH CANCER NETWORK EXPERIENCE

You have accessJournal of UrologyBladder Cancer: Superficial II1 Apr 20121781 WHAT IS THE RISK OF HIGH RISK NON-MUSCLE INVASIVE BLADDER CANCER? AN ENGLISH CANCER NETWORK EXPERIENCE Omar Al Kadhi, Robert Mills, Mark Rochester, Alexandra Colquhoun, David Neal, and William Turner Omar Al KadhiOmar Al Kadhi Cambridge, United Kingdom More articles by this author , Robert MillsRobert Mills Norwich, United Kingdom More articles by this author , Mark RochesterMark Rochester Norwich, United Kingdom More articles by this author , Alexandra ColquhounAlexandra Colquhoun Cambridge, United Kingdom More articles by this author , David NealDavid Neal Cambridge, United Kingdom More articles by this author , and William TurnerWilliam Turner Cambridge, United Kingdom More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.1798AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES High risk non-muscle invasive bladder cancer (HRNMIBC) carries a high risk of progression, and a decreased probability of survival. There is often an understandable reluctance on behalf of patients and treating urologists to submit to early radical cystectomy (RC), in favor of intravesical Bacillus Calmette-Guérin (BCG) treatment. With the advent of centralization of cancer care in English cancer networks, all RC are now performed in cancer centers. We assessed understaging of HRNMIBC in all RC done in two cancer networks between January 2008 and September 2011 avoiding the selection bias inherent in previously published series. METHODS Data on all patients undergoing RC performed in two tertiary referral centers were retrospectively reviewed. We determined the rate of muscle invasion in patients undergoing RC as first treatment (primary RC) for HRNMIBC and also in patients undergoing RC for BCG failure. We also examined the outcomes in patients with and without pre-cystectomy bladder carcinoma in situ (Tis). RESULTS A total of 120 RC procedures for presumed HRNMIBC were carried out in the two centers during the 45 month period. There were 103 men. The median age was 70 years (range 46-85). Primary RC was done in 47 patients, and RC was done in 73 patients who had failed intravesical BCG. Ten of 47 (21%) patients in the primary RC group and 19 of 73 (26%) patients in the BCG failure group were upstaged to at least pT2 disease. In the primary RC group, six patients (13%) had Tis; none were upstaged at cystectomy. In contrast, 32 patients with Tis underwent RC after failed BCG treatment (44%) and in 10 out of 32 (31%) muscle invasion was evident following RC. CONCLUSIONS In our comprehensive unselected series from two English cancer networks 1 in 5 patients who had primary RC for HRNMIBC had unsuspected muscle invasive bladder cancer; in patients who failed BCG, 1 in 4 had unsuspected muscle invasion. The highest risk of upstaging across this whole cohort was in those patients who had failed BCG and who had Tis. All patients with newly diagnosed HRNMIBC should be seen by a urologist with expertise in RC and advised that they should at least consider RC because of their high risk of unsuspected muscle invasive cancer. Patients who opt for BCG but fail, should see a cystectomy specialist promptly, because their risk of unsuspected muscle invasive cancer maybe even higher. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e719 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Omar Al Kadhi Cambridge, United Kingdom More articles by this author Robert Mills Norwich, United Kingdom More articles by this author Mark Rochester Norwich, United Kingdom More articles by this author Alexandra Colquhoun Cambridge, United Kingdom More articles by this author David Neal Cambridge, United Kingdom More articles by this author William Turner Cambridge, United Kingdom More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Pathological responses to gemcitabine/platinum-based neoadjuvant chemotherapy for muscle-invasive urothelial cancer

Pathological responses to gemcitabine/platinum-based neoadjuvant chemotherapy for muscle-invasive urothelial cancer

Risk-adapted screening in bladder cancer using the open-access Internet-based questionnaire RiskCheck bladder cancer: First evaluation by the health services research foundation IQUO, Germany.

263 Background: Till this day urologists are waiting on symptomatic persons to initiate any diagnostic work-up to identify bladder cancer (BC) patients. In result we diagnose a quarter to a third of our patients as muscle-invasive cancers. The open-access questionnaire RiskCheck bladder cancer (RCBC) was proven in a pilot-study in daily routine from German urologists organized in the health services research foundation IQUO on asymptomatic patients. Methods: The open-access RCBC questionnaire was used in urological offices to check asymptomatic patients for their BC risk exposure (personal, smoking, occupation and medical induced). The tool delivers the classical risk stratification in low- intermediate- and high risk. All people with intermediate and high risk were checked for tumor presence by urine diagnostics and in case of suspect results controlled by cystoscopy. Statistical analysis was made by IBM-SPSS 19 for incidence distribution and correlation between risk stratification and tumor detection was proven by classification tree analysis, significance p &lt; 0.05. Results: Out of 196 checked asymptomatic persons 185 (93.4%) were negative for tumor and 11 had a detectable tumor. In the group of NED 125 (68.1%) persons were classified as low risk, 26 (15.7%) as intermediate and 30 (16.2%) as high risk. Out of the 11 detected tumors 9 were at intermediate or high risk (81.8%). This resulted in an over all detection rate of 5.6% and focused on the risk population of 13.2%. The association of tumor presence and increased risk was significant (p &lt; 0.01). Compared to the western incidence rates this is an increase in effectiveness of 377. Conclusions: Risk-adapted screening in bladder cancer delivers a reasonable approach to diagnose bladder cancer before emerging symptoms. The questionnaire RCBC integrates evidence based bladder cancer inductors, is easy in use and as a open-access tool available in 10 languages via the Internet ( www.riskcheck-bladder-cancer.info) .

Radical cystectomy for clinically muscle invasive bladder cancer: does prior non-invasive disease affect clinical outcomes?

To compare clinical and pathologic outcomes of radical cystectomy for muscle invasive bladder cancer in relation to prior history of non-invasive urothelial carcinoma. Retrospective data collected from 1,150 patients managed by radical cystectomy for urothelial carcinoma of the bladder from the Canadian Bladder Cancer Network were analysed. Patients with clinical stage T2 or more were included and divided into two groups: (Group 1) patients with prior history of non-invasive urothelial carcinoma (N = 365) and (Group 2) patients with clinical muscle invasive cancer de novo (N = 785). Variables analysed included patient age, gender, pathologic stage, adjuvant chemotherapy, recurrence and mortality. Both groups were nearly equal in mean age and gender distribution, with mean ages of 67.2 and 66.7 years, and 79.7 and 79.5%, respectively (P = 0.4 and 0.9, respectively). The presence of preoperative hydronephrosis was 20.8 and 32.6% (P = 0.0007) for groups 1 and 2, respectively. The rate of higher pathological stage (T3 or T4) was 36.3 and 58% (P < 0.0001), positive lymph nodes were 20.1 and 28.8% (P = 0.002), and lymphovascular invasion was 31.7 and 46.2% (P = 0.0001) for groups 1 and 2, respectively. The rate of adjuvant chemotherapy was 15.5 and 23.3% (P = 0.002) for groups 1 and 2, respectively. None of the sampled patients received neoadjuvant chemotherapy. The overall survival (OS) and disease-specific survival (DSS) rates at 5 years were 62 and 70% for group 1 and 51 and 60% for group 2, respectively, while at 10 years, OS and DSS were 46 and 66% for group 1 and 35 and 49% for group 2, respectively (P = 0.0001 and 0.0002, respectively). Using multivariate analysis examining factors affecting recurrence and survival, we found that previous non-invasive bladder tumour history was associated with a significantly reduced risk of mortality and recurrence (Hazard ratio of 0.7 for all risks, P = 0.0002). Our retrospective study suggests that patients with non-invasive urothelial carcinoma of the bladder that progress to muscle invasion and require radical cystectomy appear to have better pathologic and clinical outcome than patients presenting with clinical muscle invasive disease de novo.

Clinical study of second transurethral resection combined with instillation therapy and transfusion therapy of dendritic cells pulsed with tumor cells in non muscle-invasive bladder cancer

Objective To observe the clinical efficacy and safety of second transurethral resection combined with instillation therapy and transfusion therapy of dendritic cells pulsed with tumor cells on non muscle-invasive bladder cancer.Methods Eighty patients with stage T1 non muscle-invasive bladder cancer were included in this protocol in which all patients prospectively received second transurethral resection within 4 to 6 weeks following initial resection.All 80 cases were divided into a DC group and a control group.In the DC group,dendritic cells pulsed with tumor cells were transfused between 6 -8 weeks.Bladder instillation therapy and follow-up was applied on the control group.The recurrence rate,the clinical efficacy and adverse reactions were observed and compared between the two groups.Results In the initial resection,21.3％,67.5％ and 11.2％ had G1,G2 and G3 transitional cell carcinoma,respectively.Twenty-seven (33.7％) had residual tumors at the second TUR,8 patients had Ta(29.6％ ) and 19 had T1 (70.4％).After the initial TUR-Bt,residual tumors were detected in 11.1％,70.4％ and 18.5％ in G1,G2 and G3,respectively.In the 8 Ta cases,2 cases moved to a higher grade,while the grade was unchanged in 6 cases.In the 19 cases with stage T1,12 had a higher grade,5 had a lower grade and 2 remained the same.In the DC group,5 cases suffered chills and fever when dendritic cells were transfused.The fever was releaved when dexamethasone was administered.The white blood cells count,creatinine and alanine aminotransferase had no statistically significance change at pre-therapy,one year after therapy and two years after therapy (P ＞0.05).The index of CD4 、CD8 、CD4/CD8 had statistically significance change at pre-therapy,one year after therapy and two years after therapy ( P ＜ 0.05 ),while the difference between one year after therapy and two years after therapy was not statistically significance ( P ＞ 0.05 ).The first and second year recurrence rate was 2％ and 6％ in the DC group,while in the control group it was 20％ and 30％.The difference was statistically significant ( P ＜ 0.05 ).Conclusion Second transurethral resection combined with instillation therapy and transfusion therapy of dendritic cells pulsed with tumor cells could be an effective therapeutic approach to lower the recurrence rate on non muscle-invasive bladder cancer. Key words: Non muscle-invasive bladder urothelium cancer; Second transurethral resection; Dendritic cells

Use of Restaging Bladder Tumor Resection for Bladder Cancer Among Medicare Beneficiaries

To evaluate the use and effectiveness of restaging bladder tumor resection using population-based data. Restaging bladder tumor resection improves staging accuracy and the response to intravesical therapy. However, its use outside of a tertiary care setting, and its subsequent clinical implications, are unknown. We identified 62 016 patients diagnosed with bladder cancer between 1992 and 2005 using SEER-Medicare data. Restaging bladder tumor resection was defined as 2 or more resections occurring within 60 days of diagnosis. Using multivariable models, we assessed the relationship between the use of restaging resection and cancer-specific survival. Restaging resection was performed in only 3064 (4.9%) of newly diagnosed bladder cancer patients, but was most common among those with high grade (7.7% vs 2.0% in low grade, P < .001) and stage (8.8% in T2 vs 2.8% in Ta/Tis, P < .001) disease. Compared to patients with muscle-invasive cancers who did not undergo restaging at diagnosis, restaging resection was associated with improved 5-year cancer-specific mortality among pathologically staged patients (20.4% vs 28.0%, P = .02), while clinically staged patients trended toward improved mortality (28.2% vs 31.9%, P = .07). Restaging transurethral resection for bladder cancer is relatively uncommon and associated with improved survival among patients with muscle invasive bladder cancer. Greater use of restaging warrants further investigation as a simple means of improving outcomes among patients suspected of having muscle invasive disease.

Potential Role of CCND1 G870A Genotype as a Predictor for Urothelial Carcinoma Susceptibility and Muscle-Invasiveness in Taiwan

The cell cycle regulator cyclin D1 (CCND1) is thought to play a major role in the transition of cell cycle from G1 to S phase. It is known that a common CCND1 G870A genotype is associated with bladder cancer in Japan and China, but not in the Western World. There is neither a report about its role in Taiwan's population, nor its genetic role of CCND1 G870A in another worldwide urothelial cancer, upper tract urothelial cancer (UTUC). Therefore, we aimed at investigating the role of CCND1 G870A in both bladder cancer and UTUC in Taiwanese cohorts. The CCND1 G870A genotypes of 171 (101 bladder cancer and 70 UTUC) patients and 243 control subjects were determined by PCR-RFLP and their correlation with clinical and histopathological data was evaluated. The genotype analysis results showed that CCND1 GG genotype was associated with a lower risk overall in urothelial (P = 0.008, OR = 0.44, 95% CI = 0.24-0.81) and bladder cancer patients (P = 0.008, OR = 0.34, 95% CI = 0.15-0.76) than those of the AA genotype. In addition, patients carrying the AG genotype had a 0.29-fold lower odds ratio of muscle-invasive cancer procession (95% CI = 0.12-0.70) compared with those carrying the AA genotype in bladder cancer patients. Surprisingly, the GG genotype had a 5.88-fold higher odds ratio of muscle-invasive cancer procession (95% CI = 1.08-32.01) compared with those carrying the AA genotype in UTUC. No association between any CCND1 G870A genotype and higher-grade risk was found. Our results suggest that the G allele of the CCND1 G870A polymorphism may be a potential predictive and prognostic biomarker to distinguish between bladder cancer and UTUC in Taiwan.

Activation of the PI3K/Akt/mTOR pathway correlates with tumour progression and reduced survival in patients with urothelial carcinoma of the urinary bladder

Phosphatidylinositol 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) pathway dysregulation has been implicated in the development of urothelial carcinoma. However, its clinical relevance has not been substantially validated in human samples. The aim of this study was to assess the expression of the pathway in a large cohort of bladder cancers using the tissue microarray technique. Immunohistochemical stains for phosphatase and tensin homologue (PTEN), phosphorylated Akt, mTOR, S6 and 4E-BP1 were performed for 887 cases, and the results were correlated with clinicopathological characteristics. The high expression of p-S6 and p-Akt corresponded significantly with high-grade and advanced-stage, while losses of PTEN and p-4E-BP1 were observed more often in high-grade and high-stage tumours. High expression of p-Akt and p-S6 predicted progression and cancer-specific mortality for non-muscle-invasive cancers treated by transurethral resection, and p-Akt was an independent factor in multivariate analysis. High expression of p-mTOR and p-Akt correlated with higher cumulative incidence of cancer-specific mortality for muscle-invasive cancer, and p-mTOR was an independent prognostic factor. We have demonstrated the impact of PI3K/Akt/mTOR alteration on the biological behaviour of bladder tumours. Proper immunohistochemical examination of the PI3K/Akt/mTOR pathway can provide useful prognostic information, and the findings may represent an additional therapeutic avenue in the treatment of bladder cancers.

Abstract 2361: The role of thrombomodulin in bladder cancer

Abstract Bladder cancer is one of the most common cancers in the United States and Taiwan. We have discovered that thrombomodulin (TM) is up-regulated in bladder cancer patients and cancer cell lines but not in normal cells. TM is an exclusive endothelial cell surface glycoprotein and an important vascular protective molecule that has anticoagulant, anti-inflammatory and anti-apoptotic properties. TM immunohistochemical analysis of tissue microarray from bladder cancer patients of the United States was performed, and we found that TM staining was more pronounced in high grade and muscle invasive cancers. TM expression by western blot have been examined in 10 bladder cancer tissues from Taiwan in comparison to 3 normal bladder tissues, 80% (8/10) of the cancer group were over-expressed while none showed in normal bladder tissues. In a cell model, we knocked down the TM expression in BFTC905 bladder cancer cells (which was a cell line from invasive TCC) by siRNA. The two stable TM knocked-down cells (C6 and E2) showed a lower proliferation rate than BFTC905 wild type cells and vector control cells (D3). TM knocked-down cells (C6 and E2) also had lower migration and invasion abilities than control cells. Furthermore, TM knocked-down cells have a decreased colony number and size in tumor formation assays. By contrast, SV-HUC-1(immortalized human normal urothelial cell line) enhanced the migration and invasion ability by transient over-expression with TM plasmid. In an animal model, the RT-PCR analysis of the ascites from mice with peritoneal inoculation of MBT-2 cells (mouse bladder tumor cell line), we found that TM expression has higher in the group with tumor formation than the group without. These findings suggest that TM may be involved in the tumor progression of bladder cancer. Thus TM could be a bladder cancer prognosis biomarker and also be a therapeutic target. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2361. doi:10.1158/1538-7445.AM2011-2361

Cyclin-Dependent Kinase-Associated Protein Cks2 is Associated with Bladder Cancer Progression

In this observational retrospective study, expression of possible cancer-related genes was measured in patients with a pathological diagnosis of superficial bladder cancer. Further measurements were made in those who subsequently developed muscle-invasive cancer. Seven of the 45 patients with superficial bladder cancer progressed to muscle-invasive cancer. Expression of fatty acid binding protein 5 (FABP5), poly(A) binding protein cytoplasmic 1 (PABPC1), DEAD box polypeptide 5 (DDX5), splicing factor 3b subunit 1 (SF3B1), murine mammary tumour integration site 6 (EIF3S6), tropomyosin 2β (TPM2), transgelin (TAGLN) and cyclin-dependent kinase-associated protein (Cks2) genes was measured in bladder samples using real-time reverse transcription-polymerase chain reaction. FABP5, PABPC1, DDX5, SF3B1, EIF3S6 and Cks2 expression levels were significantly increased, and TPM2 and TAGLN were significantly decreased, in superficial bladder cancer compared with normal bladder tissue. In patients who developed muscle-invasive cancer, the Cks2 gene showed significantly increased expression after, compared with before, invasion. The Cks2 gene may have potential as a biomarker for predicting superficial bladder cancer progression to muscle-invasive cancer.