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Environmental Health Perspectives Mar 1983N-Butyl-N-(4-hydroxybutyl)nitrosamine (BBN) is a potent carcinogen in the urinary bladder of animals. The BBN model of bladder cancer is an excellent model of human...
N-Butyl-N-(4-hydroxybutyl)nitrosamine (BBN) is a potent carcinogen in the urinary bladder of animals. The BBN model of bladder cancer is an excellent model of human urinary bladder cancer and has already led to a greater knowledge of its pathogenesis. In our studies, histogenesis and morphological characteristics of BBN urinary bladder cancer were analyzed in different animal species such as rats, mice, hamsters and guinea pigs and also in different rat strains. Papillary or nodular hyperplasia (PN hyperplasia) is found to be a preneoplastic lesion of the rat urinary bladder. Therefore, the promoting and inhibitory effects of various chemicals in two-stage urinary bladder carcinogenesis were judged by measuring PN hyperplasia in rats. Dose-dependent and organ-specific effects of the urinary bladder promoter, saccharin, in the induction of PN hyperplasia were shown in rats after initiation by BBN. The promoting effect of saccharin was seen more clearly in the urinary bladder of rats after potent initiation. A strain difference in susceptibility of the urinary bladder to the promoter was also shown. These results suggest that the above various factors may also have modifying activities on urinary bladder carcinogenesis in man.
Topics: Animals; Butylhydroxybutylnitrosamine; Carcinogens; Cell Transformation, Neoplastic; Neoplasms, Experimental; Nitrosamines; Rats; Saccharin; Tryptophan; Urinary Bladder Neoplasms
PubMed: 6832095
DOI: 10.1289/ehp.8349217 -
PloS One 2021Bladder cancer, one of the most prevalent malignancies worldwide, remains hard to classify due to a staggering molecular complexity. Despite a plethora of diagnostic...
Single-cell RNAseq and longitudinal proteomic analysis of a novel semi-spontaneous urothelial cancer model reveals tumor cell heterogeneity and pretumoral urine protein alterations.
Bladder cancer, one of the most prevalent malignancies worldwide, remains hard to classify due to a staggering molecular complexity. Despite a plethora of diagnostic tools and therapies, it is hard to outline the key steps leading up to the transition from high-risk non-muscle-invasive bladder cancer (NMIBC) to muscle-invasive bladder cancer (MIBC). Carcinogen-induced murine models can recapitulate urothelial carcinogenesis and natural anti-tumor immunity. Herein, we have developed and profiled a novel model of progressive NMIBC based on 10 weeks of OH-BBN exposure in hepatocyte growth factor/cyclin dependent kinase 4 (R24C) (Hgf-Cdk4R24C) mice. The profiling of the model was performed by histology grading, single cell transcriptomic and proteomic analysis, while the derivation of a tumorigenic cell line was validated and used to assess in vivo anti-tumor effects in response to immunotherapy. Established NMIBC was present in females at 10 weeks post OH-BBN exposure while neoplasia was not as advanced in male mice, however all mice progressed to MIBC. Single cell RNA sequencing analysis revealed an intratumoral heterogeneity also described in the human disease trajectory. Moreover, although immune activation biomarkers were elevated in urine during carcinogen exposure, anti-programmed cell death protein 1 (anti-PD1) monotherapy did not prevent tumor progression. Furthermore, anti-PD1 immunotherapy did not control the growth of subcutaneous tumors formed by the newly derived urothelial cancer cell line. However, treatment with CpG-oligodeoxynucleotides (ODN) significantly decreased tumor volume, but only in females. In conclusion, the molecular map of this novel preclinical model of bladder cancer provides an opportunity to further investigate pharmacological therapies ahead with regards to both targeted drugs and immunotherapies to improve the strategies of how we should tackle the heterogeneous tumor microenvironment in urothelial bladder cancer to improve responses rates in the clinic.
Topics: Animals; Butylhydroxybutylnitrosamine; Carcinogens; Cell Line, Tumor; Disease Models, Animal; Female; Humans; Longitudinal Studies; Male; Mice; Mice, Inbred C57BL; Polymerase Chain Reaction; Proteinuria; Proteomics; Sequence Analysis, RNA; Single-Cell Analysis; Urinary Bladder; Urinary Bladder Calculi; Urothelium
PubMed: 34232958
DOI: 10.1371/journal.pone.0253178 -
PloS One 2017Intravesical bacillus Calmette-Guerin (BCG) treatment is the most common therapy to prevent progression and recurrence of non-muscle invasive bladder cancer (NMIBC)....
Intravesical bacillus Calmette-Guerin (BCG) treatment is the most common therapy to prevent progression and recurrence of non-muscle invasive bladder cancer (NMIBC). Although the immunoreaction elicited by BCG treatment is well documented, those induced by intravesical treatment with chemotherapeutic agents are much less known. We investigated the immunological profiles caused by mitomycin C, gemcitabine, adriamycin and docetaxel in the N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN)-induced orthotopic bladder cancer mouse model. Ninety mice bearing orthotopic bladder cancer induced by BBN were randomly divided into six groups and treated with chemotherapeutic agents once a week for four weeks. After last treatment, bladder and serum samples were analyzed for cell surface and immunological markers (CD4, CD8, CD56, CD204, Foxp3, and PD-L1) using immunohistochemistry staining. Serum and urine cytokine levels were evaluated by ELISA. All chemotherapeutic agents presented anti-tumor properties similar to those of BCG. These included changes in immune cells that resulted in fewer M2 macrophages and regulatory T cells around tumors. This result was compatible with those in human samples. Intravesical chemotherapy also induced systemic changes in cytokines, especially urinary interleukin (IL)-17A and granulocyte colony stimulating factor (G-CSF), as well as in the distribution of blood neutrophils, lymphocytes, and monocytes. Our findings suggest that intravesical treatment with mitomycin C and adriamycin suppresses protumoral immunity while enhancing anti-tumor immunity, possibly through the action of specific cytokines. A better understanding of the immunoreaction induced by chemotherapeutic agents can lead to improved outcomes and fewer side effects in intravesical chemotherapy against NMIBC.
Topics: Adjuvants, Immunologic; Administration, Intravesical; Animals; Antineoplastic Combined Chemotherapy Protocols; Butylhydroxybutylnitrosamine; Cytokines; Deoxycytidine; Doxorubicin; Female; Gene Expression Regulation, Neoplastic; Humans; Mice; Mitomycin; Neoplasms, Experimental; Treatment Outcome; Urinary Bladder Neoplasms; Gemcitabine
PubMed: 28406993
DOI: 10.1371/journal.pone.0175494 -
The Journal of Pathology Nov 2018Recent studies of muscle-invasive bladder cancer show that FGFR3 mutations are generally found in a luminal papillary tumour subtype that is characterised by better... (Comparative Study)
Comparative Study
Recent studies of muscle-invasive bladder cancer show that FGFR3 mutations are generally found in a luminal papillary tumour subtype that is characterised by better survival than other molecular subtypes. To better understand the role of FGFR3 in invasive bladder cancer, we examined the process of tumour development induced by the tobacco carcinogen OH-BBN in genetically engineered models that express mutationally activated FGFR3 S249C or FGFR3 K644E in the urothelium. Both occurrence and progression of OH-BBN-driven tumours were increased in the presence of an S249C mutation compared to wild-type control mice. Interestingly, at an early tumour initiation stage, the acute inflammatory response in OH-BBN-treated bladders was suppressed in the presence of an S249C mutation. However, at later stages of tumour progression, increased inflammation was observed in S249C tumours, long after the carcinogen administration had ceased. Early-phase neutrophil depletion using an anti-Ly6G monoclonal antibody resulted in an increased neutrophil-to-lymphocyte ratio at later stages of pathogenesis, indicative of enhanced tumour pathogenesis, which supports the hypothesis that suppression of acute inflammation could play a causative role. Statistical analyses of correlation showed that while initial bladder phenotypes in morphology and inflammation were FGFR3-dependent, increased levels of inflammation were associated with tumour progression at the later stage. This study provides a novel insight into the tumour-promoting effect of FGFR3 mutations via regulation of inflammation at the pre-tumour stage in the bladder. Copyright © 2018 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Topics: Animals; Butylhydroxybutylnitrosamine; Cell Transformation, Neoplastic; Cholecystitis, Acute; Disease Models, Animal; Disease Progression; Female; Genetic Predisposition to Disease; Lymphocytes; Male; Mice, Inbred C57BL; Mice, Transgenic; Mutation; Neutrophil Infiltration; Neutrophils; Phenotype; Receptor, Fibroblast Growth Factor, Type 3; Time Factors; Tumor Microenvironment; Urinary Bladder; Urinary Bladder Neoplasms; Urothelium
PubMed: 30043421
DOI: 10.1002/path.5143 -
In Vivo (Athens, Greece) 2017This study aimed to evaluate the utility of several biological parameters for the prediction of tumor development and animal welfare in a rat model of urinary bladder...
BACKGROUND/AIM
This study aimed to evaluate the utility of several biological parameters for the prediction of tumor development and animal welfare in a rat model of urinary bladder cancer.
MATERIALS AND METHODS
The control group (n=9) received tap water while the test group (n=12) received the carcinogen N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) in drinking water. A score sheet with biological variables was used to monitor animals' welfare. Body weight, food and drink consumption and rectal temperature were measured weekly. Blood and urine samples were collected.
RESULTS
Animals from the control group exhibited a slightly higher body weight and body weight gain. The final urine volume was higher in BBN group (p<0.05). All animals from the BBN group exhibited macroscopic hematuria at 35th week. Four animals were anemic in the last week of the experiment.
CONCLUSION
The routine control of hematuria was a useful non-invasive biomarker of disease progression that may be used as a potential earlier humane endpoint. Animals did not show clinical signs of suffering that justified their sacrifice before the end of the study.
Topics: Animals; Body Weight; Butylhydroxybutylnitrosamine; Carcinogenesis; Carcinogens; Disease Models, Animal; Endpoint Determination; Humans; Rats; Urinary Bladder; Urinary Bladder Neoplasms
PubMed: 29102928
DOI: 10.21873/invivo.11172 -
Bladder Cancer (Amsterdam, Netherlands) Oct 2015Bladder cancer is one of the most common malignant genitourinary diseases worldwide. Despite advances in surgical technique, medical oncology and radiation therapy,...
Bladder cancer is one of the most common malignant genitourinary diseases worldwide. Despite advances in surgical technique, medical oncology and radiation therapy, cure of invasive tumors remains elusive for patients with late stage disease. Therefore, new therapeutic strategies are needed to improve the response rates with regard to recurrence, invasion and metastasis. Inhibitor of DNA binding (Id) proteins have been proposed as therapeutic targets due to the key regulatory role they exert in multiple steps of cancer. We aimed to explore the role of Id proteins in bladder cancer development and the pattern of expression of Id proteins in bladder carcinomas. We used a well-established chemically induced model of bladder carcinogenesis. Wild type and Id-deficient mice were given N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) in the drinking water and urinary bladder lesions were analyzed histopathologically and stained for Id1. We assessed the effects of Id1 inactivation in cultured bladder cancer cells and in a model of metastatic lung colonization. We also performed Id1 staining of human urothelial carcinoma samples and matched lymph node metastases. Id1 protein was overexpressed in the BBN-induced model of bladder cancer. Id1 deficiency resulted in the development of urinary bladder tumors with areas of extensive hemorrhage and decreased invasiveness when compared to wild type mice. Id1 inactivation led to decreased cell growth and lung colonization of human bladder cancer cells. Immunohistochemistry performed on human urothelial carcinoma samples showed Id1 positive staining in both primary tumors and lymph node metastases. In summary, our studies reveal the physiological relevance of Id1 in bladder cancer progression and suggest that targeting Id1 may be important in the development of novel therapies for the treatment of bladder cancer.
PubMed: 27376116
DOI: 10.3233/BLC-150023 -
International Journal of Nanomedicine 2017Bladder-sparing options are being developed for muscle-invasive bladder cancer in place of radical cystectomy, including the combination of chemotherapy and radiation...
Bladder-sparing options are being developed for muscle-invasive bladder cancer in place of radical cystectomy, including the combination of chemotherapy and radiation therapy. We reasoned that improving the radiotherapy component of chemoradiation could improve the control of locally advanced disease. Previously, we showed that gold nanoparticles (AuNPs) are potent enhancers of radiation therapy. We hypothesized that if AuNPs were to preferentially localize to bladder tumors, they may be used to enhance the radiation component of muscle-invasive bladder tumor therapy. Mice were treated with the carcinogen N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) for 17, 20, and 22 weeks - long enough to induce muscle-invasive tumors. Mice were then anesthetized and injected intravenously with 1.9 nm AuNPs of which most were rapidly cleared from the blood and excreted after a 30-50 minute residence time in the bladder. We found AuNPs distributed throughout the bladder wall, but most of the AuNPs were associated with the stroma surrounding the tumor cells or extracellular keratin produced by the tumor cells. There were relatively few AuNPs in the tumor cells themselves. The AuNPs therefore localized to tumor-associated stroma and this tumor specificity might be useful for specific X-ray dose enhancement therapy of muscle-invasive bladder carcinomas.
Topics: Animals; Butylhydroxybutylnitrosamine; Carcinogens; Gold; Metal Nanoparticles; Mice, Inbred C57BL; Muscles; Tissue Distribution; Urinary Bladder Neoplasms
PubMed: 29138560
DOI: 10.2147/IJN.S140977 -
Oncotarget May 2016Recent studies showed the potential linkage of estrogen/estrogen receptor signaling with bladder tumorigenesis, yet detailed mechanisms remain elusive. Here we found a...
Recent studies showed the potential linkage of estrogen/estrogen receptor signaling with bladder tumorigenesis, yet detailed mechanisms remain elusive. Here we found a new potential therapy with the combination of Bacillus Calmette-Guerin (BCG) and the anti-estrogen ICI 182,780 led to better suppression of bladder cancer (BCa) than BCG alone. Mechanism dissection found ICI 182,780 could promote BCG attachment/internalization to the BCa cells through increased integrin-α5β1 expression and IL-6 release, which may enhance BCG-induced suppression of BCa cell growth via recruiting more monocytes/macrophages to BCa cells and increased TNF-α release. Consistently, in vivo studies found ICI 182,780 could potentiate the anti-BCa effects of BCG in the carcinogen-induced mouse BCa models. Together, these in vitro and in vivo results suggest that combining BCG with anti-estrogen may become a new therapeutic approach with better efficacy to suppress BCa progression and recurrence.
Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; BCG Vaccine; Butylhydroxybutylnitrosamine; Cell Line, Tumor; Cell Movement; Cell Proliferation; Drug Synergism; Estradiol; Estrogen Receptor Antagonists; Estrogen Receptor alpha; Female; Fulvestrant; Humans; Integrin alpha5beta1; Interleukin-6; Macrophages; Mice; Neoplasms, Experimental; Treatment Outcome; Tumor Necrosis Factor-alpha; Urinary Bladder Neoplasms
PubMed: 27092883
DOI: 10.18632/oncotarget.8756 -
Oncology Reports Jun 2008The chemopreventive efficacy of cranberry juice concentrate in an experimental model of urinary bladder cancer was evaluated using female Fischer-344 rats. The animals...
The chemopreventive efficacy of cranberry juice concentrate in an experimental model of urinary bladder cancer was evaluated using female Fischer-344 rats. The animals received N-butyl-N-(4-hydroxybutyl)-nitrosamine (OH-BBN) for a period of eight weeks. Cranberry juice concentrate was administered at doses of 1.0 or 0.5 ml/rat/day beginning one week after the final OH-BBN treatment and continuing until the end of the study. The urinary bladders of all the rats were weighed and examined grossly for lesions, and all masses were submitted for pathological evaluation. A dose-dependent preventive effect of cranberry treatment was observed, with a reduced number of urinary bladder cancers (38%) in the 1.0 ml/rat/day group versus the control group. The cranberry extract neither affected body weight gain nor caused other signs of toxicity. For the metabolic studies, serum and urine were collected at 4 and 12 h after the administration of the cranberry juice concentrate and were analyzed by LC-MS/MS. Quercetin and its methylated derivative were detected in the urine samples. However, no quercetin was detected in the serum samples, indicating its poor bioavailability. These data suggest that components of cranberries may be effective in preventing urinary bladder carcinogenesis.
Topics: Animals; Butylhydroxybutylnitrosamine; Chromatography, Liquid; Female; Phytotherapy; Plant Extracts; Rats; Rats, Inbred F344; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Urinary Bladder Neoplasms; Vaccinium macrocarpon
PubMed: 18497966
DOI: No ID Found -
Cancer Prevention Research... Mar 2020Daily dosing of either NSAIDs or EGFR inhibitors has been shown to prevent bladder cancer development in a N-butyl-(4-hydroxybutyl)nitrosamine (OH-BBN)-induced rat...
Daily dosing of either NSAIDs or EGFR inhibitors has been shown to prevent bladder cancer development in a N-butyl-(4-hydroxybutyl)nitrosamine (OH-BBN)-induced rat model. However, these inhibitors cause gastrointestinal ulceration and acneiform rash, respectively, limiting their continuous use in a clinical prevention setting. We studied chemopreventive efficacy of pulsatile dosing of EGFR inhibitor erlotinib (42 mg/kg BW, once/week) combined with intermittent or continuous low doses of the NSAID naproxen (30 mg/kg BW/day, 3 weeks on/off or 128 ppm daily in diet) in the OH-BBN induced rat bladder cancer model. The interventions were started either at 1 or 4 weeks (early intervention) or 3 months (delayed intervention) after the last OH-BBN treatment, by which time the rats had developed microscopic bladder lesions. All combination regimens tested as early versus late intervention led to the reduction of the average bladder tumor weights (54%-82%; < 0.01 to < 0.0001), a decrease in tumor multiplicity (65%-85%; < 0.01 to < 0.0001), and a decrease in the number of rats with large palpable tumors (>200 mg; 83%-90%; < 0.01 to < 0.0001). Levels of signal transduction markers, Ki-67, cyclin D1, IL1β, pSTAT3, and pERK, were significantly ( < 0.05 to < 0.001) reduced in the treated tumors, demonstrating their potential utility as predictive markers for efficacy. These findings demonstrate that significant chemopreventive efficacy could be achieved with alternative intervention regimens designed to reduce the toxicity of agents, and that starting erlotinib and/or naproxen treatments at the time microscopic tumors were present still conferred the efficacy.
Topics: Animals; Anticarcinogenic Agents; Butylhydroxybutylnitrosamine; Carcinogens; Disease Progression; Drug Administration Schedule; Drug Therapy, Combination; ErbB Receptors; Erlotinib Hydrochloride; Female; Humans; Naproxen; Neoplasm Recurrence, Local; Neoplasms, Experimental; Pulse Therapy, Drug; Rats; Time Factors; Time-to-Treatment; Urinary Bladder; Urinary Bladder Neoplasms
PubMed: 31818850
DOI: 10.1158/1940-6207.CAPR-19-0339