Survival In Bladder Cancer Patients Research Articles

The Prognostic Value of Previous Irradiation on Survival of Bladder Cancer Patients.

Background: Radiation exposure is an established risk factor for bladder cancer, however consensus is lacking on the survival characteristics of bladder cancer patients with a history of radiation therapy (RT). Confounding patient comorbidities and baseline characteristics hinders prior attempts at developing such a consensus. Objective: To compare the survival characteristics of patients with suspected radiation-induced second primary cancer (RISPC) of the bladder to those with de novo bladder cancer, taking into account the patient comorbidities and baseline characteristics predictive of survival. Methods: Retrospective analysis of patients with muscle-invasive (≥T2a) or BCG-refractory stage Tis-T1 urothelial bladder cancer. Patients were excluded if prior RT exposure was used as treatment for bladder cancer or if cause of death was due to post-operative complications. A digit matching propensity score algorithm was used to match patients with prior radiation treatment to those without prior treatment. Cox regression analysis for time until death was performed following creation of the propensity score matched sample. Results: 29 patients with history of RT were matched with two controls each, resulting in a dataset of 87 observations in the event model. Results from the Cox model indicate a significantly increased hazard ratio for death at 2.22 (p = 0.047, 95% CI: 1.015–4.860) given a history of prior radiation therapy. Conclusions: In a small cohort, bladder cancer patients who underwent cystectomy had a significantly higher risk of death in the face of prior pelvic RT. This effect was found to be independent of surgical complications, numerous established patient characteristics and comorbidities traditionally predictive of survival.

Highlights of This Issue

The bone is a preferred site for metastatic dissemination and homing for many cancers. In this study, Gutierrez-Uzquiza and colleagues identified PKCϵ, a protein kinase C isoform that is highly expressed in prostate cancer, as a novel player in skeletal metastasis. Silencing PKCϵ in PC3-ML cells, an androgen-independent cell model system with high bone tropism, severely impaired the formation of bone metastatic foci in mice. Furthermore, PKCϵ is required for prostate cancer cell invasiveness and transendothelial cell migration (TEM). This study provides experimental evidence for targeting PKCϵ for the treatment of bone metastatic disease.Expression and function of the vascular endothelial growth factor receptor 1 (VEGFR1) in epithelial tumor cells has been well documented. In the current article, Ye and colleagues identified pseudogene FLT1P1 transcripts that are highly homologous to VEGFR1 mRNA expressed in colorectal cancer cells. Mechanistic studies showed that FLT1P1 noncoding transcripts play a vital role in translational regulation of cognate gene VEGFR1 expression, as well as modulation of noncognate gene expression through interaction with microRNAs. Finally, inhibiting FLT1P1 significantly impacted cell and tumor growth. These findings provide a putative target for non-coding RNA intervention in colorectal cancer.Tantamount to the identification of genetic mutations in patient tumors is the need to identify those mutations that drive tumor growth. From known bladder cancer mutations, Hensel and colleagues bioinformatically predicted which mutations promote loss of function. Then a screen was designed to mirror loss of function by introducing a shRNA library into cells and injecting the cells into mice. Genomic sequencing of the tumors revealed a number of dominate shRNAs and subsequent validation studies confirmed that loss of IQGAP1 leads to a dramatic increase in tumorigenic properties in vitro and in vivo. Importantly, reduced expression of IQGAP1 correlates with decreased bladder cancer patient survival indicating that this directed screen uncovered a novel tumor growth suppressor with biomarker potential in bladder cancer.Cancers bearing KRAS mutations are common, providing ample numbers of cases to suggest that specific mutant KRAS isoforms have distinct biological behaviors; the KRAS G13D mutation in colon cancer is a noteworthy example. Biochemical and structural analyses show that KRAS G13D rapidly exchanges its bound nucleotide, relative to other isoforms, due to a destabilizing interaction between the codon 13 aspartate side-chain and the phosphate portion of the ligand. These crystallographic insights from G13D and other KRAS mutants combined with the biochemical properties such as rates of intrinsic or GAP-stimulated GTP hydrolysis and the strength of protein-protein interactions between mutant KRAS isoforms and their effectors provides a framework for subclassifying KRAS driven cancers for better clinical decision making.

GOLPH3 is a potential therapeutic target and a prognostic indicator of poor survival in bladder cancer treated by cystectomy.

Golgi phosphoprotein 3 (GOLPH3) has been reported to be involved in the development of several human cancers. However, its clinical significance and biological role in bladder cancer remains unclear. In this study, we sought to analyze the GOLPH3 expression in bladder cancer samples and cells, and explore its clinical significance and biological role. We found that GOLPH3 was significantly increased in bladder cancer tissues and cells. Overexpression of GOLPH3 had significant correlation with poorer survival for bladder cancer patients treated by cystectomy. Knockdown of GOLPH3 inhibited the proliferation, migration and invasion of cancer cells, and tumor growth in a xenograft mouse model. GOLPH3 silencing inhibited AKT/m-TOR signaling, increased the cyclin-dependent kinase (CDK) inhibitor p27 and decreased the CDK regulator cyclin D1 and matrix metallopeptidase 9 (MMP9). Thus, GOLPH3 is likely to play important roles in bladder cancer progression via modulating AKT/mTOR signaling, and it is a novel prognostic biomarker and promising therapeutic target for bladder cancer.

Abstract 3066: Regulation of oncogenic Zeb1 by microRNA-23b in bladder cancer

Abstract MicroRNAs (miRNAs) are small, non-coding RNAs that regulate gene expression by targeted repression of transcription and translation. In this study we show that miRNA-23b (miR-23b) acts as a tumor suppressor in bladder cancer. Quantitative real-time PCR analysis showed that miR-23b is significantly down-regulated in bladder cancer cell lines and tumor tissues compared to non-malignant cells and normal tissue samples. We also demonstrate that miR-23b expression has a potential to be diagnostic and prognostic biomarker in bladder cancer. High miR-23b expression is positively correlated with higher overall survival of bladder cancer patients as revealed by Kaplan-Meier analysis. ROC analysis showed that miR-23b expression can distinguish between normal and bladder cancer tissues. Further we elucidated the biological significance of miR-23b in bladder cancer. Over-expression of miR-23b in bladder cancer cells inhibited cell proliferation and impaired colony formation. Fluorescence activated cell sorting (FACS) analysis revealed that re-expression of miR-23b in bladder cancer cells induced G0/G1 cell cycle arrest and apoptosis while inhibiting cell migration and invasion. Luciferase reporter assays demonstrated that Zeb1, a crucial regulator of epithelial-to-mesenchymal transition (EMT), is a direct target of miR-23b in bladder cancer. These results show that loss of miR-23b confers a proliferative advantage and promotes bladder cancer cell migration and invasion. Furthermore, re-expression of miR-23b may be a beneficial therapeutic strategy for the treatment of human bladder cancer. Citation Format: Shahana Majid, Altaf A. Dar, Sharanjot Saini, Soichiro Yamamura, Varahram Shahryari, Yozo Mitsui, Yuichiro Tanaka, Rajvir Dahiya, Guoren Deng. Regulation of oncogenic Zeb1 by microRNA-23b in bladder cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3066. doi:10.1158/1538-7445.AM2015-3066

The c-MET Network as Novel Prognostic Marker for Predicting Bladder Cancer Patients with an Increased Risk of Developing Aggressive Disease

Previous studies have shown that c-MET is overexpressed in cases of aggressive bladder cancer (BCa). Identification of crosstalk between c-MET and other RTKs such as AXL and PDGFR suggest that c-MET network genes (c-MET-AXL-PDGFR) may be clinically relevant to BCa. Here, we examine whether expression of c-MET network genes can be used to identify BCa patients at increased risk of developing aggressive disease. In vitro analysis, c-MET knockdown suppressed cell proliferation, invasion, and migration, and increased sensitivity to cisplatin-induced apoptosis. In addition, c-MET network gene (c-MET, AXL, and PDGFR) expression allowed discrimination of BCa tissues from normal control tissues and appeared to predict poor disease progression in non-muscle invasive BCa patients and poor overall survival in muscle invasive BCa patients. These results suggest that c-MET network gene expression is a novel prognostic marker for predicting which BCa patients have an increased risk of developing aggressive disease. These genes might be a useful marker for co-targeting therapy, and are expected to play an important role in improving both response to treatment and survival of BCa patients.

Tumour-suppressive microRNA-144-5p directly targets CCNE1/2 as potential prognostic markers in bladder cancer.

Background:Analysis of a microRNA (miRNA) expression signature of bladder cancer (BC) by deep-sequencing revealed that clustered miRNAs microRNA (miR)-451a, miR-144-3p, and miR-144-5p were significantly downregulated in BC tissues. We hypothesised that these miRNAs function as tumour suppressors in BC. The aim of this study was to investigate the functional roles of these miRNAs and their modulation of cancer networks in BC cells.Methods:The functional studies of BC cells were performed using transfection of mature miRNAs. Genome-wide gene expression analysis, in silico analysis, and dual-luciferase reporter assays were applied to identify miRNA targets. The association between miR-144-5p levels and expression of the target genes was determined, and overall patient survival as a function of target gene expression was estimated by the Kaplan–Meier method.Results:Gain-of-function studies showed that miR-144-5p significantly inhibited cell proliferation by BC cells. Four cell cycle-related genes (CCNE1, CCNE2, CDC25A, and PKMYT1) were identified as direct targets of miR-144-5p. The patients with high CCNE1 or CCNE2 expression had lower overall survival probabilities than those with low expression (P=0.025 and P=0.032).Conclusion:miR-144-5p functions as tumour suppressor in BC cells. CCNE1 and CCNE2 were directly regulated by miR-144-5p and might be good prognostic markers for survival of BC patients.

The Impact of Perioperative Blood Transfusion on Survival of Bladder Cancer Patients Submitted to Radical Cystectomy: Role of Anemia Status

BackgroundThe prognostic role of perioperative blood transfusion (PBT) in patients who underwent radical cystectomy (RC) for bladder cancer (BCa), although supported by clinical evidence, still remains to be assessed definitively. ObjectiveTo investigate the impact of PBT on RC patients for overall survival and after stratifying according to preoperative anemia status and to define whether the oncologic impact may be assumed to be a primary effect of PBT or attributed to the reduced preoperative hemoglobin (Hb) level. Design, setting, and participantsA total of 1490 consecutive patients with nonmetastatic BCa who underwent RC and pelvic lymph node dissection between January 1990 and August 2013 at a single referral center entered the study. PBT and preoperative Hb levels were statistically correlated with postoperative oncologic outcomes. Outcome measurements and statistical analysisKaplan-Meier analyses were used to evaluate the impact of PBT on overall mortality (OM) and cancer-specific mortality (CSM). Multivariable Cox regression analyses tested the relationship between PBT and the risk of OM and CSM in the overall population and after stratifying patients according to the severity of their anemia. Results and limitationsA total of 580 patients (38.9%) received PBT. Mean postoperative follow-up was 125.13 mo (median: 110 mo). Overall 5- and 10-yr CSM survival rates were 58.3% and 47.6%, respectively. At multivariable Cox regression analyses, PBT could not be associated with an increased risk of either CSM or OM (all p > 0.3). Conversely, preoperative Hb levels were significantly associated with OM (hazard ratio [HR]: 0.88; confidence interval [CI], 0.83–0.95) and CSM (HR: 0.84; 95% CI, 0.77–0.95) (all p<0.001). A significant detrimental effect of PBT on OM (HR: 1.65; 95% CI, 1.08–2.52) and CSM (HR: 1.68; 95% CI, 1.04–2.70) (all p<0.03) was found in patients without preoperative anemia status. ConclusionsIn nonanemic BCa patients proposed for RC, PBT is associated with a significant detrimental effect on CSM and OM. This effect seems to be attributable to the direct impact of PBT, regardless of the preoperative Hb value. Patient summaryIn nonanemic patients proposed for radical cystectomy, a negative prognostic impact of perioperative blood transfusion should be taken into account.

Uncovering the clinical utility of miR-143, miR-145 and miR-224 for predicting the survival of bladder cancer patients following treatment

Accurate prognosis is a key factor in establishing optimal therapeutic decisions; yet in the case of bladder cancer (BlCa) current prognostic indicators cannot ensure optimal disease management. Here, we aimed to evaluate the previously unexplored clinical potential of the urological cancer-related miR-145, miR-143 and miR-224 in BlCa. A total of 279 bladder tissue specimens were included in this study (133 BlCa, 107 adjacent normal and 39 healthy samples). Total RNA was extracted from tissues, it was polyadenylated and reverse transcribed to cDNA. The expression of target molecules was measured via quantitative real-time PCR. The expression levels of both miR-143 and miR-145 were significantly decreased, whereas those of miR-224 were increased in BlCa. Receiver operating characteristic curve analysis indicated a significant discriminatory capacity for miR-143/miR-145 levels. Important associations with disease aggressiveness were observed for all three microRNAs; elevated levels were observed in tumors of higher stage and grade, as well as in 'high-risk' TaT1 patients. More importantly, high miR-143/145 levels could effectively prognose inferior overall survival for muscle-invasive patients and could independently predict the progression of superficial tumors. Finally, the combination of miR-143/145 overexpression with the widely used prognostic markers of European Organization for Research and Treatment of Cancer-risk groups or recurrence at the first follow-up cystoscopy resulted to a superior positive prediction of non-muscle-invasive bladder cancer short-term progression compared with the use of the abovementioned markers alone. The cancer-related miR-143, miR-145 and miR-224 were investigated for the first time in the clinical setting of BlCa, and miR-143/145 cluster constitutes a novel marker helpful for providing an enhanced prediction of oncologic outcome for BlCa patients.

Metabonomic profiling of bladder cancer.

Early diagnosis and life-long surveillance are clinically important to improve the long-term survival of bladder cancer patients. Currently, a noninvasive biomarker that is as sensitive and specific as cystoscopy in detecting bladder tumors is lacking. Metabonomics is a complementary approach for identifying perturbed metabolic pathways in bladder cancer. Significant progress has been made using modern metabonomic techniques to characterize and distinguish bladder cancer patients from control subjects, identify marker metabolites, and shed insights on the disease biology and potential therapeutic targets. With its rapid development, metabonomics has the potential to impact the clinical management of bladder cancer patients in the future by revolutionizing the diagnosis and life-long surveillance strategies and stratifying patients for diagnostic, surgical, and therapeutic clinical trials. An introduction to metabonomics, typical metabonomic workflow, and critical evaluation of metabonomic investigations in identifying biomarkers for the diagnosis of bladder cancer are presented.

Prognostic Value of 11C-Choline PET/CT and CT for Predicting Survival of Bladder Cancer Patients Treated with Radical Cystectomy

Background: In patients with bladder cancer (BCa) preoperative staging with ¹¹C-choline positron emission tomography-computed tomography (PET/CT) could be used to derive prognostic information and hence stratify patients preoperatively with respect to disease management. Methods: From June 2004 to May 2007, 44 patients with localized BCa were staged with ¹¹C-choline PET/CT before radical cystectomy. The results of imaging were correlated to overall survival (OS) and cumulative incidence of cancer-specific death (CSD). Results: There was no statistically significant difference in OS and CSD between the patient groups when stratified for organ-confined versus non-organ-confined disease or lymph node involvement defined by either ¹¹C-choline PET/CT (OS: p = 0.262, hazard ratio [HR] = 1.60; p = 0.527, HR = 0.76; CSD: p = 0.144, HR = 2.25; p = 0.976, HR = 0.98) or CT (OS: p = 0.518, HR = 1.34; p = 0.228, HR = 1.67; CSD: p = 0.323, HR = 1.90; p = 0.136, HR = 2.38). The limitation of this study is the small number of included patients. Conclusion: In our prospective trial neither CT nor ¹¹C-choline PET/CT were able to sufficiently predict OS or CSD in BCa patients treated with radical cystectomy albeit trends and moderately increased HRs could be demonstrated without significant differences between CT or ¹¹C-choline PET/CT. However, these trends might prove statistically significant in bigger patient cohorts. Therefore initial transsectional imaging might be of clinical relevance in respect to prognosis and could play a role in the counseling of BCa patients.

Prognostic significance of the epithelial-to-mesenchymal transition markers e-cadherin, vimentin and twist in bladder cancer.

The goal of this study was to utilize long-term patient follow-up to determine whether epithelial-to-mesenchymal transition (EMT)-related markers can predict bladder cancer patient survival and progression of disease. This study included 121 patients with bladder cancer. Sixty-four of these patients presented with non-muscle invasive (NMI, stage T1) bladder cancer and 57 with muscle invasive (MI, stage T2, T3). The patients were diagnosed and treated between May 1998 and July 2012. The EMT markers E-cadherin, Twist, and Vimentin were detected via immunohistochemistry. Univariate and multivariate/Cox analyses were then utilized to determine whether these EMT markers could be useful prognostic markers for predicting bladder cancer patient outcomes. Analysis of the 121 bladder cancer patients in this study revealed that the frequency of E-cadherin expression was 59.5% (72/121), Twist was 54.5% (66/121), and Vimentin was 24.8% (30/121). Twist and Vimentin were found to have statistically significant correlations with grade, recurrence, and progression but not with stage, whereas E-cadherin was associated with stage but not with the other parameters. In the univariate analysis, grade (p = 0.02) was the only significant predictor for progression-free survival (PFS). Stage, grade, and expression of E-cadherin, Vimentin and Twist were included in the multivariate analysis of predicting PFS. In this analysis, grade (p = 0.01) and Vimentin expression (p = 0.001) were found to be significant prognostic factors in predicting PFS. Grade and Vimentin are potential independent indicators in predicting bladder cancer progression and survival.

Incidence and survival analysis of renal cell carcinoma patients among permanent residents in Yangpu district of Shanghai during 2002-2012

Objective To analyze the incidence and mortality of bladder cancer patients among permanent residents in Yangpu district of Shanghai from January 2002to December 2012.Methods The data of bladder cancer patients among the permanent residents of Yangpu district were obtained from the database of cancer registration and management system in Shanghai.The temporal trend in the incidence and mortality of bladder cancer was assessed using annual percent change(APC) model.Kaplan-Meier analysis with log-rank test was performed for survival analysis.The incidence and mortality were standardized with age composition of standard population from a nationwide census carried out in 2000.Results A total of 1 252cases with bladder cancer were newly diagnosed between 2002and 2012in Yangpu district,and it accounted for 2.69%ofthe total patients with malignant diseases.The annual crude incidence of bladder cancer was 10.51/105,with that of males being significantly higher than that of females(15.59/105 vs 5.18/105,P0.01).The standardized incidence was 4.83/105, with that of males being also significantly higher than that of females(7.26/105 vs 2.32/105,P0.01).The crude incidence of bladder cancer showed a significant increase(APC=4.14,P0.01)from 2002to 2012;however,the standardized incidence showed no significant increase.The incidence increased rapidly after 50years old.A total of 690cases died of bladder cancer during the study period.The annual crude mortality of bladder cancer was 8.61/105 in males,which was significantly higher than that in females(2.84/105,P0.01).The standardized martality was 3.64/105 in males and 1.15/105 in females(P 0.01).The crude mortality of bladder cancer were increased significantly and the standardized one did not alter significantly during this period.The 5-year survival rate of the newly diagnosed bladder cancer patients was 63.15%,with that of surgically treated ones being significantly higher that of non-surgically treated ones(74.31% vs 51.80%,P0.01). Conclusion The crude incidence and mortality of bladder cancer are greatly increased during 2002-2012.Surgical treatment can significantly prolong the survival of bladder cancer patients.Early diagnosis and prompt surgical resection can improve the overall survival of bladder cancer patients.

Urine tenascin-C is an independent risk factor for bladder cancer patients

Urine biomarkers offer a non‑invasive method of detecting bladder cancer, monitoring disease progression and predicting disease recurrence and therapeutic treatment efficacy. Tenascin‑C (TN‑C), as a component of the extracellular matrix, is vital in the progression of bladder cancer. However, there is little to report with regard to urine TN‑C and its correlation with bladder cancer grade, stage, recurrence and prognosis. In the present study, 66 samples of voided urine from patients with bladder cancer and 42 samples from volunteers were obtained. The urine TN‑C concentration was determined using an ELISA assay. The correlation between the urine TN‑C concentration and the tumor grade, stage and time from bladder cancer diagnosis to recurrence was analyzed by a rank correlation analysis. Multivariate Cox proportional hazards regression was used for finding the main life‑threatening factors among age, gender, tumor grade, stage, relapse and the urine TN‑C concentration. At the end, the Kaplan‑Meier method was used to evaluate the survival rate affected by urine TN‑C as a single factor. The results indicated that the urine TN‑C concentration in the bladder cancer patients was higher compared with the healthy control volunteers (22.5 times higher). Among all the patients, urine TN‑C concentration had a positive correlation with the bladder cancer grade and stage, with correlation coefficients of 0.905 and 0.308, respectively; however, this correlation was negative between urine TN‑C concentration and the time from bladder cancer diagnosis to recurrence. Moreover, the multivariate Cox proportional hazards model analysis indicated that urine TN‑C, like tumor grade and recurrence, may be an independent risk factor for bladder cancer patient survival. However, it is noteworthy that inflammation may affect the concentration of urine TN‑C. The results of the present study indicate that urine TN‑C may be used as a biomarker for monitoring the recurrence of bladder cancer in patients and for predicting its prognosis. However, inflammation of the urinary tract should be excluded first.

The HER4 isoform JM-a/CYT2 relates to improved survival in bladder cancer patients but only if the estrogen receptor α is not expressed

Bladder cancer tumors expressing human epidermal growth factor receptor 4 (HER4) demonstrate improved patient survival. HER4 isoforms and estrogen receptor alpha (ER-α) can form chaperone complexes causing cell-proliferation. We wanted to explore if HER4 isoforms and ER-α could correlate to poor prognosis in bladder cancers. We developed mRNA assays for HER4 isoforms (JM-a, JM-b, CYT1, and CYT2) and for ER-α. Expression was analyzed in tumors from 85 bladder cancer patients and compared to overall survival (median follow-up of 5.1 years). ER-α was expressed in 38% (n = 32) of tumors but did not correlate to survival (p = 0.4698). HER4 was expressed in 42% (n = 36) and in all cases as the ER-α binding isoform JM-a. The JM-a isoform can be alternatively spliced to either a CYT1 isoform (JM-a/CYT1) or a CYT2 isoform (JM-a/CYT2). All HER4 expressing tumors expressed the JM-a/CYT2 isoform and half of those (18/36) expressed both isoforms. JM-a/CYT2 expression correlated to improved survival (p = 0.004), but not when ER-α was co-expressed (p = 0.897). Immunohistochemistry revealed protein expression of HER4 and ER-α in tumor cells. Growth of RT4 bladder cancer cells, expressing both JM-a/CYT2 and ER-α was inhibited by the specific ER-α inhibitor raloxifene. Likewise, stable transfection with JM-a/CYT2 inhibited the growth of T24 bladder cancer cells, but only when ER-α was inhibited. Our results demonstrate that HER4 JM-a/CYT2 expressing bladder cancers relate to favorable prognosis when ER-α is not co-expressed. In vitro studies indicate that ER-α inhibition may be a useful treatment for patients with tumors expressing both ER-α and HER4 JM-a/CYT2.

MicroRNA-23b Functions as a Tumor Suppressor by Regulating Zeb1 in Bladder Cancer

MicroRNAs (miRNAs) are small, non-coding RNAs that regulate gene expression by targeted repression of transcription and translation. In this study we show that miRNA-23b (miR-23b) acts as a tumor suppressor in bladder cancer. Quantitative real-time PCR analysis showed that miR-23b is significantly down-regulated in bladder cancer cell lines and tumor tissues compared to non-malignant cells and normal tissue samples. We also demonstrate that miR-23b expression has a potential to be diagnostic and prognostic biomarker in bladder cancer. High miR-23b expression is positively correlated with higher overall survival of bladder cancer patients as revealed by Kaplan-Meier analysis. ROC analysis showed that miR-23b expression can distinguish between normal and bladder cancer tissues. Further we elucidated the biological significance of miR-23b in bladder cancer. Over-expression of miR-23b in bladder cancer cells inhibited cell proliferation and impaired colony formation. Fluorescence activated cell sorting (FACS) analysis revealed that re-expression of miR-23b in bladder cancer cells induced G0/G1 cell cycle arrest and apoptosis while inhibiting cell migration and invasion. Luciferase reporter assays demonstrated that Zeb1, a crucial regulator of epithelial-to-mesenchymal transition (EMT), is a direct target of miR-23b in bladder cancer. These results show that loss of miR-23b confers a proliferative advantage and promotes bladder cancer cell migration and invasion. Furthermore, re-expression of miR-23b may be a beneficial therapeutic strategy for the treatment of human bladder cancer.

Overexpression of Rab25 contributes to metastasis of bladder cancer through induction of epithelial–mesenchymal transition and activation of Akt/GSK-3β/Snail signaling

Rab25, an epithelial-specific member of the Rab family of small guanosine triphosphatases, is associated with several human cancers. The goal of this study was to determine its function in bladder cancer (BC). We examined the Rab25 expression pattern in two different cohorts of BC patients treated with radical cystectomy by quantitative PCR, western blotting and immunohistochemical staining. A series of in vitro and in vivo assays were performed to elucidate the function of Rab25 in BC and its underlying mechanisms. Rab25 expression was significantly elevated at both the messenger RNA and protein levels in BCs compared with normal bladder tissues. High Rab25 expression was closely associated with lymph node (LN) metastasis and was an independent predictor for poor disease-free survival in BC patients. Downregulation of Rab25 in BC cells markedly inhibited invasive motility in vitro and metastatic potential in vivo. In addition, downregulation of Rab25 in BC EJ and T24 cells increased the expression levels of epithelial markers (E-cadherin and α-catenin) and decreased the levels of mechamechy markers (vimentin and fibronectin). Simultaneously, downregulation of Rab25 in EJ and T24 cells resulted in the inactivation of downstream phosphorylated protein kinase B (p-Akt), phosphorylated glycogen synthase kinase-β (p-GSK-3β) and snail signaling. This study demonstrates that Rab25 can promote BC metastasis through induction of epithelial-mesenchymal transition process and activation of Akt/GSK-3β/Snail signaling pathway; Rab25 expression level can predict LN metastasis and inferior clinical outcome in BC patients.

Polymorphisms in theXRCC1gene modify survival of bladder cancer patients treated with chemotherapy

Survival of bladder cancer patients depends on several factors including disease stage and grade at diagnosis, age, health status of the patient and the applied treatment. Several studies investigated the role of DNA repair genetic variants in cancer susceptibility, but only few studies investigated their role in survival and response to chemotherapy for bladder cancer. We genotyped 28 single nucleotide polymorphisms (SNP) in DNA repair genes in 456 bladder cancer patients, reconstructed haplotypes and calculated a score for combinations of the SNPs. We estimated Hazard Ratios (adjHR) for time to death. Among patients treated with chemotherapy, variant alleles of five SNPs in the XRCC1 gene conferred better survival (rs915927 adjHR 0.55 (95%CI 0.32-0.94); rs76507 adjHR 0.48 (95%CI 0.27-0.84); rs2854501 adjHR 0.25 (95%CI 0.12-0.52); rs2854509 adjHR 0.21 (95%CI 0.09-0.46); rs3213255 adjHR 0.46 (95%CI 0.26-0.80). In this group of patients, an increasing number of variant alleles in a XRCC1 gene score were associated with a better survival (26% decrease of risk of death for each additional variant allele in XRCC1). By functional analyses we demonstrated that the previous XRCC1 SNPs confer lower DNA repair capacity. This may support the hypothesis that survival in these patients may be modulated by the different DNA repair capacity determined by genetic variants. Chemotherapy treated cancer patients bearing an increasing number of "risky" alleles in XRCC1 gene had a better survival, suggesting that a proficient DNA repair may result in resistance to therapy and shorter survival. This finding may have clinical implications for the choice of therapy.

Abstract 3095: Oncogenic miRNA-182-5p targets Smad4 and RECK in human bladder cancer.

Abstract Abstract Introduction and Objective: Onco-miR-182-5p has been reported to be over-expressed in bladder cancer (BC) tissues however a detailed functional analysis of miR-182-5p has not been carried out in BC. Therefore the purpose of this study was to: 1. conduct a functional analysis of miR-182-5p in bladder cancer, 2. assess its usefulness as a tumor marker, 3. identify miR-182-5p target genes in BC. Methods: Initially we found that miR-182-5p expression was significantly higher in bladder cancer compared to normal tissues and high miR-182-5p expression was associated with shorter overall survival in BC patients. To study the functional significance of miR-182-5p, we over-expressed miR-182-5p with miR-182-5p precursor. Results: Cell proliferation, migration and invasion abilities were increased in BC cells. However cell apoptosis was inhibited by miR-182-5p. We also identified Smad4 and RECK as potential target genes of miR-182-5p using several algorithms. 3’UTR luciferase activity of these target genes was significantly decreased and protein expression of these target genes was significantly up-regulated in miR-182-5p inhibitor transfected bladder cancer cells. MiR-182-5p also increased nuclear beta-catenin expression and while Smad4 repressed nuclear beta-catenin expression. Conclusions: Our data suggests that miR-182-5p plays an important role as an oncogene by knocking down RECK and Smad4, resulting in activation of the Wnt-beta-catenin signaling pathway in bladder cancer. Source of Funding: Grants RO1CA138642, RO1CA130860, RO1CA160079, VA Research Enhancement Award Program (REAP) and Merit Review grants Citation Format: Hiroshi Hirata, Koji Ueno, Varahram Shahryari, Yuichiro Tanaka, Z. Laura Tabatabai, Yuji Hinoda, Rajvir Dahiya. Oncogenic miRNA-182-5p targets Smad4 and RECK in human bladder cancer. [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 3095. doi:10.1158/1538-7445.AM2013-3095

High expression of HuR in cytoplasm, but not nuclei, is associated with malignant aggressiveness and prognosis in bladder cancer.

IntroductionHuman antigen R (HuR) regulates the stability of mRNA and is associated with cell proliferation, angiogenesis, and lymphangiogenesis. However, the clinical significance and pathological role of HuR in bladder cancer remains unclear. The main objective of this investigation was to clarify the relationships between HuR expression and clinical significance and cancer cell proliferation, angiogenesis, lymphangiogenesis, and expressions of cyclooxygenase (COX)-2 and vascular endothelial growth factor (VEGF)-A, -C, and -D.MethodsAll expressions were examined by immunohistochemical techniques in 122 formalin-fixed specimens of bladder cancer patients. HuR expression was evaluated separately with cytoplasmic and nuclear staining. Cell proliferation, angiogenesis and lymphangiogenesis were measured as the percentage of Ki-67-positive cell (proliferation index, PI), CD34-stained vessels (microvessel density, MVD), and D2-40-stained vessels (lymph vessel density, LVD). Relationships between each HuR expression and clinicopathological features, prognosis, and expressions of COX-2 and VEGFs were analyzed by multi-variate analyses. HuR expression was also investigated in 10 mice of N-Butyl-N-[4-hydroxybutil] nitrosamine (BBN) induced bladder cancer model.ResultsIn human tissues, high cytoplasmic expression was seen in 5% and 25.4% of normal and cancer cells, respectively. Nuclear HuR expression bore no significant relationship to any pathological features. However, cytoplasmic HuR expression appeared positively associated with pT stage and grade (P<0.001). In mouse tissues, similar trends were confirmed. Cytoplasmic expression correlated with PI, MVD, and LVD, as well as expression of VEGF-A and -C, but not VEGF-D. High cytoplasmic expression of HuR was a significant predictor of metastasis and cause-specific survival, and was identified as a prognostic correlative factor for metastasis (hazard ratio, 4.75; P = 0.028) in a multivariate analysis model that included pathological features.ConclusionsCytoplasmic HuR appears to play important roles in cell proliferation, progression, and survival of bladder cancer patients. Its expression was associated with angiogenesis, lymphangiogenesis, and expressions of VEGF-A and –C.

ERCC1 and XRCC1 expression predicts survival in bladder cancer patients receiving combined trimodality therapy

Combined trimodality therapy, including transurethral resection and platinum-based chemoradiotherapy, has shown promising results for muscle-invasive bladder cancer. However, this type of treatment may decrease survival as a result of delayed cystectomy in patients with non-responding tumors. DNA repair proteins may affect survival of bladder cancer patients receiving combined trimodality therapy, by affecting the perioperative nature of the tumor cells or by repairing DNA damaged by platinum agents and radiation. We investigated the associations of excision repair cross-complementing group 1 (ERCC1), X-ray repair cross-complementing group 1 (XRCC1) and apurinic/apyrimidinic endonuclease 1 (APE1) expression with response and survival in 157 locally advanced bladder cancer patients receiving combined trimodality therapy, in order to determine the predictive value of the expression of these proteins in patient selection for therapy. We examined ERCC1, XRCC1 and APE1 expression in tumor specimens using immunohistochemistry. Patients positive for ERCC1, positive for XRCC1 and positive for either ERCC1 or XRCC1, exhibited significantly improved disease-specific survival rates (P=0.023, 0.025 and 0.0091, respectively). In multivariate analysis, combined ERCC1 and XRCC1 expression was independently associated with disease-specific mortality [risk ratio (RR): 0.64; 95% confidence interval (CI), 0.43-0.94 and P=0.024]. Thus, combined ERCC1 and XRCC1 expression may serve as an independent prognostic marker for survival in bladder cancer patients receiving combined trimodality therapy. Prospective studies with a larger sample size are required to confirm these results.