Development Of Urinary Bladder Cancer Research Articles

Is there a promoting role for artificial sweeteners in the evolution of bladder cancer? A meta-analysis of current literature.

Urinary bladder cancer is a frequent neoplasia in the urogenital system. Ageing and smoking are the two main risk factors, however, some chemical agents such as artificial sweeteners could act as initiators or promoters. After identifying trends in scientific literature, we conducted a wide search in PubMed database and a meta-analysis was performed on extracted data to determine the role of artificial sweeteners in the development of urinary bladder cancer. Twenty-one full reports were enrolled from screening of PubMed database into final analysis involving 116,568 subjects in comparisons. Overall, 13,682 and 102,886 cases were identified for bladder cancer patients and healthy controls, respectively. Among artificial sweetener users, 12.5% was the incidence of bladder cancer. In the control group, 11.2% of cases suffered from urothelial carcinoma of the bladder. About 40.7% of the patients suffering from urinary neoplasms and 37.8% of the healthy cases were artificial sweetener users, respectively. There were only minor differences in overall descriptive data. The incidence of urinary bladder cancer among artificial sweetener users and control cases showed no risk difference (RD: 0.00, CI: -0.06 to 0.06). The frequency of artificial sweetener use among patients suffering from urinary bladder neoplasms and healthy subjects was compared which showed equal occurrences (OR: 0.96, CI: 0.79 to 1.17). According to our results, the carcinogenic risk of artificial sweeteners is not proven. Saccharin should not be kept as a promoter in urothelial malignant transformation.

Long-term ambient hydrocarbon exposure and incidence of urinary bladder cancer

Particulate matter and volatile organic compounds, including total hydrocarbons (THCs), are major ambient air pollutants. Primary nonmethane hydrocarbons (NMHCs) originate from vehicle emissions. The association between air pollution and urinary bladder cancer (UBC) is debatable. We investigated whether long-term exposure to ambient hydrocarbons increases UBC risk among people aged ≥ 20 years in Taiwan. Linkage dataset research with longitudinal design was conducted among 589,135 initially cancer-free individuals during 2000–2013; 12 airborne pollutants were identified. Several Cox models considering potential confounders were employed. The study outcomes were invasive or in situ UBC incidence over time. The targeted pollutant concentration was divided into three tertiles: T1/T2/T3. The mean age of individuals at risk was 42.5 (SD 15.7), and 50.5% of the individuals were men. The mean daily average over 10 years of airborne THC concentration was 2.25 ppm (SD 0.13), and NMHC was 0.29 ppm (SD 0.09). Both pollutants show long-term monotonic downward trend over time using the Mann–Kendall test. There was a dose-dependent increase in UBC at follow-up. UBC incidence per 100,000 enrollees according to T1/T2/T3 exposure to THC was 60.9, 221.2, and 651.8, respectively; it was 170.0/349.5/426.7 per 100,000 enrollees, corresponding to T1/T2/T3 exposure to NMHC, respectively. Without controlling for confounding air pollutants, the adjusted hazard ratio (adj.HR) was 1.83 (95% CI 1.75–1.91) per 0.13-ppm increase in THC; after controlling for PM2.5, adj.HR was even higher at 2.09 (95% CI 1.99–2.19). The adj.HR was 1.37 (95% CI 1.32–1.43) per 0.09-ppm increase in ambient NMHC concentration. After controlling for SO2 and CH4, the adj.HR was 1.10 (95% CI 1.06–1.15). Sensitivity analyses showed that UBC development risk was not sex-specific or influenced by diabetes status. Long-term exposure to THC and NMHC may be a risk factor for UBC development. Acknowledging pollutant sources can inform risk management strategies.

Abstract 5130: Prognostic significance of loss of Cathepsin L expression in urinary bladder cancer progression

Abstract During 2021 there will be 83,730 new cases of urinary bladder cancer (UBC) in the US and 17,200 UBC deaths. Most UBC arise from the urothelial lining. UBC frequently recurs and is the most expensive cancer to treat due to the need for constant patient monitoring. New prognostic markers are needed to assist patient management. This study aims to determine if Cathepsin L is an independent prognostic factor after controlling for UBC Stage and Grade. We studied the interrelationship of Cathepsin L with Ki67, a prognostic marker in UBC. A clearer understanding of the biological mechanisms underlying UBC development and progression will lead to improved methods of UBC prognosis and suggest new therapies. Cathepsin L is a ubiquitously expressed cysteine endopeptidase capable of degrading enzymes, receptors, and transcription factors, and of activating enzymes, receptors, biologically active peptides, and pro-hormone processing by limited proteolysis. Subcellularly located in the endosomal lysosomal compartment, Cathepsin L is catalytically active at pH 3.0 - 6.5. This is a retrospective study of archived tissues and existing deidentified medical records and tumor registry information. Archived FFPE tissues were donated to Wood Hudson Cancer Research Laboratory by St. Elizabeth Healthcare (SEHC). The study was approved by the SEHC IRB. Tumors were from 41 women and 92 men. UBC were classified according to subtype, depth of invasion and grade by a Board Certified pathologist (LED). 36/62 (58.1%) of Stage 0 non-invasive UBC were Grade 1 or 2, and 26/62 (41.9%) were Grade 3 or 4. Similarly 39/90 papillary tumors (43.3%) were Grade 1 or 2 and 51/90 (36.7%) were Grade 3 or 4. In contrast 68/71 invasive UBC (Stages II, III and IV) were Grade 3 or 4. 39/90 papillary tumors were Grade 1 or 2. All non-papillary tumors were Grade 3 or 4. Ki 67 (clone MIB-1, Dako) and Cathepsin L (Abcam) expression were detected immunohistochemically. Cathepsin L was highly expressed in normal urothelium (p < 0.0001). In Stage 0, Cathepsin L expression in Grade 1,2,3 UBC was similar compared to expression in normal urothelium (p = 0.9070, 0.2560 and 0.0722). In Stage 0, Cathepsin L expression in Grade 4 UBC was significantly reduced compared to normal urothelium (p = 0.0011). Cathepsin L expression was significantly reduced in Grade 3 and 4 papillary tumors vs Grade 1 and 2 papillary tumors (p = 0.0002) and in Grade 3 and 4 non-papillary tumors vs Grade 3 and 4 papillary tumors (p = 0.0133). In each UBC subtype, grade or stage, when Cathepsin L expression was reduced cell proliferation as measured by Ki67 expression was increased and overall, this was significant (p < 0.0001). Significantly reduced Cathepsin L expression was associated with tumor recurrence and death from UBC (p = 0.03, p < 0.0001). These data suggest that when Cathepsin L is lost, intracellular levels of growth factor receptors such as EGFR may increase driving cell proliferation and UBC progression. Citation Format: Julia H. Carter, James A. Deddens, Michelle C. Robillard, Mariah K. Dooley, Daniel Stelzer, Zachary S. Taylor, Larry E. Douglass. Prognostic significance of loss of Cathepsin L expression in urinary bladder cancer progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5130.

CTSV (cathepsin V) promotes bladder cancer progression by increasing NF-κB activity

ABSTRACT Chronic inflammation is positively associated with the development of urinary bladder cancer. However, its detailed regulatory mechanism remains elusive. The quantitative real-time polymerase chain reaction was used to measure mRNA levels of relative genes. The protein levels were monitored by western blotting. Cell proliferation and viability were evaluated by the cell counting Kit 8 (CCK8) and colony formation assays, respectively. The dual-luciferase reporter assay was performed to assay the transcriptional activity. In vivo experiments were implemented in nude mice as well. The TCGA database analysis suggested that the aberrant expression of cathepsin V (CTSV) was related to a poor outcome in bladder cancer patients. CTSV boosted the inflammation reaction, which facilitated the development of bladder cancer. The overexpression of CTSV increased the proliferation and viability of bladder cancer cells. On the contrary, the deletion of CTSV significantly inhibited the proliferation and viability of bladder cancer cells. The tumor repression resulting from CTSV deficiency in vitro was also verified in vivo. Moreover, multiple cancer-associated luciferase screening showed that the overexpression of CTSV triggered the inflammatory signaling pathway, which could be restored by introducing the NF-κB inhibitor. CTSV is upregulated and promotes proliferation through the NF-κB pathway in bladder cancer and may be a potential target in inflammation-associated bladder cancer.

CircCA12 Promotes Malignant Process via Sponging miR-1184 and Upregulating RAS Family in Bladder Cancer.

Circular RNAs (circRNAs) are a panel of non-coding RNAs that mediate the regulation of gene expression, as well as pathological responses. Nonetheless, the function and expression pattern of circRNAs in urinary bladder cancer (UBC) remain unclear. Herein, we examined the function of circCA12 in UBC development. qRT-PCR results demonstrated remarkable circCA12 upregulation in UBC cell lines, as well as tissues. CCK-8, colony formation, and xenograft assays were employed to determine the effect of circCA12 on UBC. Our data illustrated silencing circCA12 repressed the proliferation along with the colony-formation capability of UBC cells. The migration and metastasis potential of UBC cells were remarkably abated in vivo, as well as in vitro after transfection with si-cirCA12 or sh-circCA12. Moreover, luciferase reporter and RIP assays indicated that circCA12 binds to miRNA-1184 through sponging miRNA, thereby up-regulating the expression of RAS family genes (NRAS, KRAS, and HRAS). In conclusion, the circCA12/miRNA-1184/RAS family was identified as a regulatory axis in UBC progression.

Association of arsenic with recurrence of urinary bladder cancer.

Arsenic is known to be an important aetiological factor for the development of urinary bladder cancer. It is known to be found excessively in ground water in certain geographical areas, including West Bengal. We have studied patients with recurrent bladder cancer from different areas of this Indian state and correlated arsenic as a causative aetiological factor for development and aggressiveness of the biological behaviour of urinary cancer. We included 31 patients from various parts of West Bengal state with recurrent bladder cancer who were operated in our institute. Their clinical and residential data and their arsenic content of tumour tissue were measured. Statistical analysis was performed to test the association of tissue arsenic with clinicopathological features of recurrent disease. We found very high levels of arsenic in tumour tissue in all residents of the districts with high prevalence of arsenic in the drinking water. We also observed more aggressive clinicopathological progression and early recurrence in patients with high arsenic content. We conclude that arsenic is a causal factor in the clinicopathological progression of recurrent urinary bladder cancer. Measures to decrease the level of arsenic in drinking water should be taken as this may both improve clinicopathological outcomes in the recurrence of urinary bladder carcinoma, as well as reducing its overall incidence.

MiADMSA ameliorate arsenic induced urinary bladder carcinogenesis in vivo and in vitro

Background and purposeArsenicosis is a major threat to public health and is a major cause of the development of urinary bladder cancer. Oxidative/ nitrosative stress is one of the key factors for these effects but the involvement of other associated factors is less known. There is a lack of data for the efficacy of chelator against urinary bladder carcinogenesis. The present study demonstrates the early signs of arsenic exposed urinary bladder carcinogenesis and its attenuation by Monoisoamyl dimercaptosuccinic acid (MiADMSA). MethodsMale rats were exposed to 50 ppm of sodium arsenite and dimethylarsinic acid (DMA) via drinking water for 18 weeks and treated with MiADMSA (50 mg/kg, orally once daily for 5 days) for 3 weeks with a gap one week between the two courses of treatments. We compared in vivo data with in vitro by co-exposing 100 nM of sodium arsenite and DMA to rat (NBT-II) as well as human transitional epithelial carcinoma (T-24) cells with 100 nM of MiADMSA. ResultsThe data showed that sodium arsenite and DMA exposure significantly increased the tissue arsenic contents, ROS, TBARS levels, catalase, SOD activities and significantly decreased GSH level which might be responsible for an increased 8−OHdG level. These changes might have increased pro-oncogenic biomarkers like MMP-9 and survivin in serum, bladder tissues, NBT-II, and T-24 cells. High cell migration and clonogenic potential in NBT-II and T-24 cells exposed to arsenic suggest pronounced carcinogenic potential. Significant recovery in these biomarkers was noted on treatment with MiADMSA. ConclusionEarly signs of urinary bladder carcinogenesis were observed in arsenic and DMA exposed rats which were linked to metal accumulation, oxidative/ nitrosative stress, 8-OHdG, MMP-9 and survivin which were reduced by MiADMSA possibly via its efficient chelation abilities in vivo and in vitro.

Downregulation of TET1 Promotes Bladder Cancer Cell Proliferation and Invasion by Reducing DNA Hydroxymethylation of AJAP1.

Ten-eleven translocation 1 (TET1) is a member of methylcytosine dioxygenase, which catalyzes 5-methylcytosine (5 mC) to 5-hydroxymethylcytosine (5 hmC) to promote the demethylation process. The dysregulated TET1 protein and 5 hmC level were reported to either suppress or promote carcinogenesis in a cancer type-dependent manner. Currently, the role of TET1 in the development of urinary bladder cancer (UBC) and its underlying molecular mechanisms remain unclear. Herein, we found that TET1 expression was downregulated in UBC specimens compared with normal urothelium and was inversely related to tumor stage and grade and overall survival, suggesting its negative association with UBC progression. TET1 silencing in UBC cells increased cell proliferation and invasiveness while the ectopic expression of wild-type TET1-CD, but not its enzymatic inactive mutant, reversed these effects and suppressed tumorigenicity in vivo. In addition, as a direct regulator of TET1 activity, vitamin C treatment increased 5 hmC level and inhibited the anchorage-independent growth and tumorigenicity of UBC cells. Furthermore, we found that TET1 maintained the hypomethylation in the promoter of the AJAP1 gene, which codes for adherens junction-associated protein 1. The downregulation of AJAP1 reversed TET1-CD-induced nuclear translocation of β-catenin, thus inhibiting the expression of its downstream genes. In human UBC specimens, AJAP1 is frequently downregulated and positively associated with TET1. Notably, low expression levels of both TET1 and AJAP1 predict poor prognosis in UBC patients. In conclusion, we found that the frequently downregulated TET1 level reduces the hydroxymethylation of AJAP1 promoter and subsequently activates β-catenin signaling to promote UBC development. The downregulation of both TET1 and AJAP1 might be a promising prognostic biomarker for UBC patients.

DNA repair gene polymorphism (XPA and XPG) and risk of urinary bladder cancer in North-Indian population

PurposeTo assess XPA A23G (rs1800975) and XPG Asp1104His (rs17655) polymorphism as risk factors for urinary bladder cancer in North-Indian population. MethodsThe prevalence of genetic polymorphisms of XPA and XPG was investigated in cases and age-gender matched healthy controls. XPA and XPG genes were genotyped using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. ResultsThe XPA GG and XPG CC genotypes were significantly associated with UBC risk (adjusted OR = 3.78, 95% CI = 1.40-10.18, p ≤ 0.05 and adjusted OR = 4.00, 95% CI = 1.19-13.43, p ≤ 0.05 respectively). After stratifying the genotypes by age, gender and smoking status, risk for carriers of XPA (AG + GG) genotype was significantly higher for males and smokers (adjusted OR = 2.55, 95% CI = 1.30–5.03, adjusted OR = 2.36, 95% CI = 1.14–4.85, p ≤0.05 respectively) while the risk for carriers of the XPG (GC + CC) genotype was significantly high for younger (median age ≤60) individuals and females (adjusted OR = 2.13, 95% CI = 1.05–4.35, adjusted OR = 18.12, 95% CI = 1.20–272.73, p ≤0.05 respectively). ConclusionXPA and XPG polymorphism might be significant risk factor in North-Indian population and acquisition of these genetic variations together with smoking might trigger urinary bladder cancer development. Further pathway based studies on larger population are warranted to confirm our conclusion. Clinical significanceThe XPA-A23G and XPG-Asp1104His genetic variations could be used as UBC susceptibility markers in North-Indian population. These SNPs could be useful biomarkers for UBC prognosis or diagnosis and thus could help in enhancing the survival rates of UBC patients and improving patient care. However, before making any further conclusion, we suggest these genetic variations should be further explored using a larger study sample for their role as potential clinical biomarkers or therapeutic targets. The role of other SNPs, gene-gene interactions and gene-environment interactions should also be taken into consideration while estimating the genetic risk of these SNPs on UBC development.

Study of risk factors for development of urinary bladder cancer in patients in Urmia; a single-center study by logit regression approach

Introduction: Urinary bladder cancer is the most common genitourinary system malignancy in humans. Objectives: Due to the high prevalence of urinary bladder cancer, having enough knowledge about urinary bladder cancer risk factors can assist in prevention and early recognition of this malignancy. Patients and Methods: In this study case-control, 109 individuals with urinary bladder tumor (as a case group) and 200 individuals without any neoplastic history aged >40 years (as a control group) were included (2016-2017). The data were collected using patients’ files, their pathology reports and also a checklist. The data were analyzed using SPSS version 20, and logistic regression test was applied to investigate the binary outcome. A P value less than 0.05 was considered significant. Results: The results showed, a significant relationship between smoking (P<0.001), having high risk occupation (P<0.001), low water intake (P<0.001) and pickle consumption (P<0.001) with urinary bladder cancer. Conclusion: Smoking is a strong risk factor for urinary bladder cancer. Our study showed smoking and other occupational carcinogens are the main causes of urinary bladder cancers. Thus preventing and controlling these risk factors are important for health schedules.

Inhibition of G9a by a small molecule inhibitor, UNC0642, induces apoptosis of human bladder cancer cells.

Urinary bladder cancer (UBC) is characterized by frequent recurrence and metastasis despite the standard chemotherapy with gemcitabine and cisplatin combination. Histone modifiers are often dysregulated in cancer development, thus they can serve as an excellent drug targets for cancer therapy. Here, we investigated whether G9a, one of the histone H3 methyltransferases, was associated with UBC development. We first analyzed clinical data from public databases and found that G9a was significantly overexpressed in UBC patients. The TCGA Provisional dataset showed that the average expression level of G9a in primary UBC samples (n = 408) was 1.6-fold as much as that in normal bladder samples (n = 19; P < 0.001). Then we used small interfering RNA to knockdown G9a in human UBC T24 and J82 cell lines in vitro, and observed that the cell viability was significantly decreased and cell apoptosis induced. Next, we choosed UNC0642, a small molecule inhibitor targeting G9a, with low cytotoxicity, and excellent in vivo pharmacokinetic properties, to test its anticancer effects against UBC cells in vitro and in vivo. Treatment with UNC0642 dose-dependently decreased the viability of T24, J82, and 5637 cells with the IC50 values of 9.85 ± 0.41, 13.15 ± 1.72, and 9.57 ± 0.37 μM, respectively. Furthermore, treatment with UNC0642 (1-20 μM) dose-dependently decreased the levels of histone H3K9me2, the downstream target of G9a, and increased apoptosis in T24 and J82 cells. In nude mice bearing J82 engrafts, administration of UNC0642 (5 mg/kg, every other day, i.p., for 6 times) exerted significant suppressive effect on tumor growth without loss of mouse body weight. Moreover, administration of UNC0642 significantly reduced Ki67 expression and increased the level of cleaved Caspase 3 and BIM protein in J82 xenografts evidenced by immunohistochemistry and western blot analysis, respectively. Taken together, our data demonstrated that G9a may be a promising therapeutic target for UBC, and an epigenetics-based therapy by UNC0642 is suggested.

Abstract 4989: Efficacy of erlotinib and/or naproxen when administered by intermittent dosing schedules in the prevention of chemically induced urinary bladder cancers

Abstract Inflammation and epidermal growth factor receptor (EGFR) signaling dysregulation plays an important role in urinary bladder cancer development. We have previously shown that single agent regimens with erlotinib (EGFR inhibitor) or naproxen (nonsteroidal anti-inflammatory drug; NSAID) when given continuously were highly effective in the prevention of OH-BBN-induced urinary bladder cancers in rats. Better tolerated chemoprevention regimens can be obtained by reducing doses and frequency of administration. Low dose combinations may also achieve substantial efficacy with minimal toxicity by targeting complementary pathways. Female Fischer-344 rats were obtained at 28 days of age, placed on Teklad (4% fat) diet, and received OH-BBN (150 mg/gavage) 2x/week for 8 weeks beginning at 56 days of age. In the first study, beginning one week after the final OH-BBN treatment the rats (25/group) received either: Group 1, erlotinib (42 mg/kg BW, 1x/week); Group 2, naproxen (30 mg/kg BW/day, 3 weeks on/ 3 weeks off); Group 3, the combination of erlotinib and naproxen (using the same treatment regimens), and Group 4, vehicle. The rats were palpated for urinary bladder tumors, weighed 1x/week and observed daily for signs of toxicity. At the end of the study (10 months after the initial OH-BBN), the average weights of the urinary bladders (bladder plus tumors combined) were determined. The weights in Groups 1-4 were: 194*, 186*, 136* and 354 mg (*P &amp;lt;0.05, Wilcoxon Rank test). This was accompanied by an increase in tumor latency, a decrease in tumor multiplicity, and a decrease in the number of rats with large palpable tumors (&amp;gt; 200 mg). The second study used the same treatment regimens, but administration of the agents was delayed until three months after the final OH-BBN treatment, at which point microscopic transitional cell carcinomas were present. At the end of the study (11 months after the initial OH-BBN), the weights of the individual bladders in Groups 1-4 were: 584, 234*, 138* and 779 mg (*P&amp;lt; 0.05, Wilcoxon Rank test). Thus, naproxen and particularly the combination were still highly effective. This was accompanied by an increase in tumor latency and a decrease in the number of rats with large palpable tumors (&amp;gt; 200 mg). These studies demonstrate that in protocols meant to reduce the toxicity of agents (weekly erlotinib or intermittent dosing with naproxen), high chemopreventive efficacy was achieved. Of importance, starting erlotinib and/or naproxen intermittent treatments at the time microscopic cancers were present still reduced the size of the urinary bladder cancers without showing observable toxicity. The latter is of particular interest based on the clinical FAP trial showing great efficacy of erlotinib and sulindac. Supported by NCI contract number HHSN261201500036I, Task Order HHSN26100002. Citation Format: Altaf Mohammed, Mark S. Miller, Ronald A. Lubet, Chen Suen, Shizuko Sei, Robert H. Shoemaker, Clinton J. Grubbs. Efficacy of erlotinib and/or naproxen when administered by intermittent dosing schedules in the prevention of chemically induced urinary bladder cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4989.

High oncogenic risk human papillomavirus and urinary bladder cancer

To determine the role of human papillomavirus (HPV) of high oncogenic risk in the development of urinary bladder cancer. 100 patients (72 men and 28 women) aged 38 to 90 years (mean age 65+/-10 years) diagnosed with bladder cancer were examined and underwent treatment. Clinical assessment was complemented by enzyme-linked immunosorbent assays for the presence of antiviral antibodies to herpes simplex virus (HSV) type 1 and type 2, cytomegalovirus (CMV), Epstein-Barr virus (EBV), urethra scraping for detecting high oncogenic risk HPV. Tumor tissue was sampled for PCR virus detection. Semi-quantitative analysis was used to evaluate the components of lymphocyte-plasmocyte and leukocyte infiltrates and cytopathic changes in tumor tissue. There were positive correlations between cytopathic cell changes (koylocytosis and intranuclear inclusions, as manifestations of HPV) and the level of antiviral antibodies, the presence of viruses in the tumor, as well as with the components of the lymphoid-plasmocyte infiltrate. Negative correlations were found between the presence of papillomatosis and the above changes. Human papillomavirus is believed to be a trigger for the initiation of a tumor in young patients with a latent infection (CMV and EBV, HSV, HPV). Cytopathic changes (kylocytosis and intranuclear inclusions) were associated with the activity and morphological features of herpes-viral infections. Their degree varied depending on the stage of the process, but not on the anaplasia degree. Papillomatosis is associated with a more favorable course of the tumor process.

Urinary cadmium levels in active and retired coal miners

ABSTRACTA meta-analysis, based upon 24 publications, showed a significantly elevated risk for urinary bladder cancer amongst miners. In European underground hard coal mining areas, an increased risk for urinary bladder cancer development was noted among hard coal miners, in particular in three investigations in the greater Dortmund area. However, the cause remains unclear. As cadmium (Cd), which was reported to be a bladder carcinogen in humans and is a constituent of coal, the aim of this study was to determine urinary Cd levels in active and retired hard coal miners and assess whether hard coal miners demonstrated elevated metal levels. In total, 103 retired and 25 active hard coal miners as well as 18 controls without any history of hard coal mining were investigated for urinary Cd levels. Urinary Cd concentrations, in addition to other elements, were analyzed in spot urines by ICP-MS-based multi-element analysis in a Department for Forensic and Clinical Toxicology. Limit of detection (LOD) for Cd was 0.5 μg/L. Reference value for occupationally non-exposed working age population was 0.8 μg/L. In total, 49% of all underground coal miners were exposed to coal dust, 12% to grinded rock, and 39% to both. Urinary Cd levels in retired as well as active coal miners and controls were clearly below the Biological Exposure Index. Urinary Cd concentration is a suitable biomarker to evaluate the metallic load of the body, as the half-life is > than 10 years. The detected urinary Cd levels in retired and active coal miners indicated underground hard coal miners were not apparently exposed to Cd to a occupationally-relevant concentration.

Transcription Factor KLF5 Binds a Cyclin E1 Polymorphic Intronic Enhancer to Confer Increased Bladder Cancer Risk.

A polymorphic KLF5 binding site near the CCNE1 gene explains genetic risk identified through GWAS. Mol Cancer Res; 14(11); 1078-86. ©2016 AACR.

Changes in Immunogenicity during the Development of Urinary Bladder Cancer: A Preliminary Study.

In the present study, we evaluated tumor-infiltrating lymphocytes (TILs) and blood regulatory T lymphocyte (Tregs, CD4+/CD25+/FoxP3+) expression in bladder cancer patients. The number of CD4+, CD8+, CD25+, FoxP3+ and CD20+ TILs was analyzed in association with clinico-pathomorphological features. In more advanced metastasizing tumors, showing non-classic differentiation (ND) and a more aggressive tissue invasion type (TIT), the number of TILs decreased. A low number of CD4+ TILs was associated with poor prognosis. Similarly, Treg frequency before surgery and after surgical treatment was significantly lower in more advanced tumors. The changes in TILs, as well as of local and systemic Tregs, were accompanied by changes in the histological phenotype of urothelial carcinoma regarding pT stage, NDs, TIT, and clinical outcomes. The number of TILs and the frequency of blood Tregs (indicators of antitumor response) may be essential for choosing an immunotherapy that is adjusted to the immune status according to the phase of tumor growth. Moreover, a significant reduction in the number of CD4+ and CD8+ TILs with the development of NDs in more advanced tumors may be associated with lower tumor immunogenicity, resulting in immune tolerance towards tumor tissue. These observations and the tendency of urothelial bladder carcinoma to undergo NDs in a heterogeneous manner during tumor progression suggest complex interactions between bladder cancer immunogenicity and stages of tumor progression.

The effects of whole green tea infusion on mouse urinary bladder chemical carcinogenesis.

Green tea (GT) is one of the most popular beverages worldwide whose beneficial effects on health have been demonstrated. Recent studies suggest that GT may contribute to reduction of cancer risk and progression. The aim of this study was to evaluate the effects of whole GT on urinary bladder chemical carcinogenesis in male and female ICR mice. The GT characterization was performed using spectrophotometric methods. Urinary bladder lesions were induced using N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) by gavage during 10 weeks and whole GT (0.5%) was provided ad libitum during 20 weeks. Animals from groups BBN+GT and BBN only developed preneoplastic lesions. We did not observe any effects by GT infusion administration on urinary bladder cancer development.

Simultaneous Activation of Kras and Inactivation of p53 Induces Soft Tissue Sarcoma and Bladder Urothelial Hyperplasia

The development of the Cre recombinase-controlled (Cre/LoxP) technique allows the manipulation of specific tumorigenic genes, temporarily and spatially. Our original intention of this study was to investigate the role of Kras and p53 in the development of urinary bladder cancer. First, to validate the effect of intravesical delivery on Cre recombination (Adeno-Cre), we examined activity and expression of β-galactosidase in the bladder of control ROSA transgenic mice. The results confirmed specific recombination as evidenced by β-galactosidase activity in the bladder urothelium of these mice. Then, we administered the same adenovirus into the bladder of double transgenic KrasLSLG12D/+. p53fl/fl mice. The virus solution was held in place by a distal urethral retention suture for 2 hours. To our surprise, there was a rapid development of a spindle-cell tumor with sarcoma characteristics near the suture site, within the pelvic area but outside the urinary track. Since we did not see any detectable β-galactosidase in the area outside of the bladder in the validating (control) experiment, we interpreted that this sarcoma formation was likely due to transduction by Adeno-Cre in the soft tissue of the suture site. To avoid the loss of skin integrity associated with the retention suture, we transitioned to an alternative technique without suture to retain the Adeno-Cre into the bladder cavity. Interestingly, although multiple Adeno-Cre treatments were applied, only urothelial hyperplasia but not carcinogenesis was observed in the subsequent experiments of up to 6 months. In conclusion, we observed that the simultaneous inactivation of p53 and activation of Kras induces quick formation of spindle-cell sarcoma in the soft tissues adjacent to the bladder but slow formation of urothelial hyperplasia inside the bladder. These results strongly suggest that the effect of oncogene regulation to produce either hyperplasia or carcinogenesis greatly depends on the tissue type.

ALKBH2, a novel AlkB homologue, contributes to human bladder cancer progression by regulating MUC1 expression

The ALKBH family of proteins are highly expressed in various types of human cancer where they are involved in tumor growth and progression. However, multiple isoforms of ALKBH exist and the effect of individual isoforms on the development of urinary bladder cancer is unknown, particularly the molecular mechanisms involved in the progression from a noninvasive to invasive phenotype. We examined the role and function of ALKBH2 in human bladder cancer development in vitro and provide the first report that suppression of ALKBH2 in a human urothelial carcinoma cell line, KU7, reduces the expression of the transmembrane mucin protein, MUC1, and induces G1 cell cycle arrest. Moreover, reduction of ALKBH2 suppressed epithelial to mesenchymal transition (EMT) via increasing E-cadherin and decreasing vimentin expression. Transfection of MUC1 siRNA inhibited cell proliferation and EMT to the same extent as ALKBH2 gene silencing in vitro. ALKBH2 knockdown significantly suppressed MUC1 expression and tumor volume of bladder cancers in vivo as assessed in an orthotopic mouse model using ALKBH2 shRNA transfected KU7 cells. Immunohistochemical examination showed high expression levels of ALKBH2 in human urothelial carcinoma samples, especially in high-grade, superficially and deeply invasive carcinomas (pT(1) and >pT(2)), and in carcinoma in situ but not in normal urothelium. This study demonstrates that ALKBH2 is an upstream molecule of the oncoprotein, MUC1, and regulates cell cycle and EMT, resulting in progression of urothelial carcinomas.

Transcriptional Modulation of the ERK1/2 MAPK and NF-kB Pathways in Human Urothelial Cells After Trivalent Arsenical Exposure: Implications for Urinary Bladder Cancer

Chronic exposure to drinking water contaminated with inorganic arsenic (iAs) is associated with an increased risk of urinary bladder (UB) cancers in humans. The exact role of specific iAs metabolite(s) in As-mediated carcinogenesis remains largely unknown. Experimental evidence suggests that trivalent arsenicals, namely arsenite (iAsIII) and two of its metabolites, monomethylarsonous acid (MMAIII) and dimethylarsinous acid (DMAIII), are possible proximate UB carcinogens. Here, we used a transcriptomics approach to examine perturbed molecular pathways in a human urothelial cell line (UROtsa) after short-term exposure to iAsIII, MMAIII and DMAIII. Molecular pathways containing genes that encode proteins implicated in UB cancer development were perturbed by both MMAIII and DMAIII. These pathways included those of the extracellular signal-regulated kinase 1/2 mitogen-activated protein kinase (ERK 1/2 MAPK) and nuclear factor kappa beta (NF-κB). Together, these results may inform the current understanding of effects in the UB induced by acute As exposure and the relationship of these effects with As-mediated carcinogenesis.