How cancer cells attach to urinary bladder epithelium in vivo: study of the early stages of tumorigenesis in an orthotopic mouse bladder tumor model (original) (raw)
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International Journal of Molecular Sciences, 2021
Non-muscle-invasive bladder cancer is the most common form of bladder cancer. The main problem in managing bladder tumors is the high recurrence after the transurethral resection of bladder tumors (TURBT). Our study aimed to examine the fate of intravesically applied cancer cells as the implantation of cancer cells after TURBT is thought to be a cause of tumor recurrence. We established an orthotopic mouse bladder tumor model with MB49-GFP cancer cells and traced them during the first three days to define their location and contacts with normal urothelial cells. Data were obtained by Western blot, immunolabeling, and light and electron microscopy. We showed that within the first two hours, applied cancer cells adhered to the traumatized epithelium by cell projections containing α3β1 integrin on their tips. Cancer cells then migrated through the epithelium and on day 3, they reached the basal lamina or even penetrated it. In established bladder tumors, E-cadherin and desmoplakin 1/2 ...
Development of a murine intravesical orthotopic human bladder cancer (mio-hBC) model
American journal of clinical and experimental urology, 2018
We have developed a murine intravesical orthotopic human bladder cancer (mio-hBC) model for the establishment of superficial urothelial cell carcinomas. In this model we catheterize female atyhmic nude mice and pre-treat the bladder with poly-L-lysine for 15 minutes, followed by intravesical instillation of luciferase-transfected human UM-UC-3 cells. Cancer cells are quantified by bioluminescent imaging which has been validated by small animal ultrasound. Poly-L-lysine pre-treatment increased engraftment rate (84.4%) and resulted in faster growing tumors than trypsin pre-treatment. In addition, tumors respond through a decrease in growth and increase in apoptosis to chemotherapy with mitomycin C. Previous intravesical models utilized KU7 cells which have been later determined to be of non-bladder origin. They display markers consistent with HeLa cells, requiring a need for a true intravesical bladder model. Efficient engraftment and rapid superficial growth patterning of the human b...
A new in vivo model to study invasion and metastasis of human bladder carcinoma
Cancer research, 1987
An animal model to investigate the invasive and metastatic properties of human bladder transitional cell carcinoma (HTCC) was established. Two long-term HTCC cell lines (RT4 and EJ) and one HTCC cell line derived in our laboratory (LD-71) were tumorigenic when injected s.c. into nude mice but had little potential to invade locally or metastasize before the animals succumbed to tumor burden. Experimental lung metastases were, however, observed in approximately 60% of animals given injections of RT4 or EJ cell lines in the tail vein. The cells were also implanted transurethrally into the urinary bladders of athymic mice. RT4 cells, which were originally isolated from a superficial papillary tumor, produced histologically noninvasive tumors after transurethral inoculation with no evidence of metastasis. In contrast EJ cells, which were originally isolated from a more aggressive tumor, produced invasive tumors in nude mouse bladders and metastasized to the lungs spontaneously. The invas...
An Orthotopic Model of Murine Bladder Cancer
Journal of Visualized Experiments, 2011
In this straightforward procedure, bladder tumors are established in female C57 mice through the use of catheterization, local cauterization, and subsequent cell adhesion. After their bladders are transurethrally catheterized and drained, animals are again catheterized to permit insertion of a platinum wire into bladders without damaging the urethra or bladder. The catheters are made of Teflon to serve as an insulator for the wire, which will conduct electrical current into the bladder to create a burn injury. An electrocautery unit is used to deliver 2.5W to the exposed end of the wire, burning away extracellular layers and providing attachment sites for carcinoma cells that are delivered in suspension to the bladder through a subsequent catheterization. Cells remain in the bladder for 90 minutes, after which the catheters are removed and the bladders allowed to drain naturally. The development of tumor is monitored via ultrasound. Specific attention is paid to the catheterization technique in the accompanying video.
Experimental biology and medicine (Maywood, N.J.), 2014
Tumor metastasis is characterized by enhanced invasiveness and migration of tumor cells through the extracellular matrix (ECM), resulting in extravasation into the blood and lymph and colonization at secondary sites. The ECM provides a physical scaffold consisting of components such as collagen fibrils, which have distinct dimensions at the nanoscale. In addition to the interaction of peptide moieties with tumor cell integrin clusters, the ECM provides a physical guide for tumor cell migration. Using nanolithography we set out to mimic the physical dimensions of collagen fibrils using lined nanotopographical silicon surfaces and to explore whether metastatic tumor cells are uniquely able to respond to these physical dimensions. Etched silicon surfaces containing nanoscale lined patterns with varying trench and ridge sizes (65-500 nm) were evaluated for their ability to distinguish between a non-metastatic (253J) and a highly metastatic (253J-BV) derivative bladder cancer cell line. ...
Histopathological characterization of a syngeneic orthotopic murine bladder cancer model
International Braz J Urol, 2008
Purpose: We developed and characterized by histopathology and immunohistochemistry a syngeneic murine bladder tumor model derived from the MB49 tumor cell line. Materials and Methods: Bladder tumor implantation was achieved by intravesical instillation of 5 x 10 5 MB49 tumor cells in C57BL/6 mice. A chemical lesion of the bladder was performed in order to promote intravesical tumor implantation. The bladder wall lesion was accomplished by transurethral instillation of silver nitrate (AgNO 3 ). After 15 days, the animals were Results: Twenty-nine out of 30 animals (96.7%) developed intravesical tumors in a 15-day period. Macroscopically, the mean bladder weight was 0.196g (0.069-0.538g), 10 to 15 times the normal bladder weight. The immunohistochemical Conclusions: cauterization is widely described in the literature for syngeneic orthotopic animal models, the technique described in this study represents an alternative for intravesical bladder tumor implantation. Moreover, the histopathology and immunohistochemical analysis of the murine bladder tumor model derived from the MB49 cell line showed a resemblance to human Key words: bladder cancer; intravesical instillation; tumor cell line; mice/c57bl; experimental neoplasm Int Braz J Urol. 2008; 34: 220-9
International Journal of Molecular Sciences
Urinary bladder cancer is often multifocal; however, the intraluminal dissemination of the urothelial cancer cells is poorly understood. The involvement of N-cadherin in the adhesion of the cancer urothelial cells to the urothelium had not previously been studied. Therefore, we herein explore the possibility of the intraluminal dissemination of the urothelial cancer cells by evaluating the role of classical cadherins in the adhesion of urothelial cancer cells to the urothelium. We used E-cadherin negative T24 cells and established a T24 Ncadlow cell line with an additionally decreased expression of N-cadherin in the plasma membrane and a decreased secretion of proform of metalloproteinase 2. The labelled T24 and T24 Ncadlow cells were seeded onto urothelial in vitro models. After 24 h in co-culture, unattached cancer cells were rinsed and urothelia with attached cancer urothelial cells were processed for fluorescence and electron microscopy. Both the T24 and T24 Ncadlow cells attach...
Animal Models for Basic and Preclinical Research in Bladder Cancer
Bladder Cancer - From Basic Science to Robotic Surgery, 2012
Bladder cancer is one of the most common cancers in the world. In 2006, there were about 61,240 diagnosed cases of bladder cancer and approximately 13,060 deaths attributable to this disease, being the prevalence estimated worldwide more than 1,000,000 patients (Jemal et al., 2006; Lerner, 2005). Taking into account that its incidence seems to be increasing, bladder cancer is clearly a significant public health issue around the world. Thus, it is necessary to intensify research on this topic. Urinary bladder cancer originates mainly from epithelial cells of the urothelium (Lopez-Beltran et al., 2004; Montironi et al., 2005). When initially diagnosed, most bladder cancers (about 70%) do not present muscle invasion, and are thus known as non-muscle invasive bladder cancer (pTa and pT1). In these cases, a simple transurethral resection is sufficient to remove the tumor. However, some patients experience recurrence or even tumor progression. The progression of the tumor involves invasion of tumor cells, which penetrate deeper layers of the bladder such as the detrusor muscle (pT2), perivesical tissue (pT3) and extravesical organs (pT4) (Figure 1). Since this progression threatens the patient's life, more aggressive therapies are necessary (Sobin et al., 1997). Intensive research in bladder cancer, as well as that in most tumors, is being carried out to elucidate the reason for the appearance of tumors, and to find out which factors are involved in their development and which are related to the tumor progression process. These investigations, which provide insights into the biology of the tumor, are essential for the implementation of new therapeutic and/or preventive modalities (Bhattacharya et al., 2010; Zhang et al., 2011). Research on basic science is focused on the mechanisms that lead cells towards transformation and development of cancer, using simple experimental models where it is easier to interpret the results. Cell culture techniques are widely used to study different oncological processes. The cell culture is the growth of any cell type, usually tumor cells, in with nutrient-containing solutions. The cells grow attached to the plastic surface, forming a monolayer, usually in a two-dimensional way. This technique allows studying processes such as mutagenesis, invasion, migration, and production of proteolytic enzymes. Although cell culture is a very important tool, it has certain limitations. Many biological processes depend on the three-dimensional architecture. In addition, monolayer culture is usually
Nanotechnology in Bladder Cancer: Diagnosis and Treatment
Cancers
Bladder cancer (BC) is the second most common cancer of the urinary tract in men and the fourth most common cancer in women, and its incidence rises with age. There are many conventional methods for diagnosis and treatment of BC. There are some current biomarkers and clinical tests for the diagnosis and treatment of BC. For example, radiotherapy combined with chemotherapy and surgical, but residual tumor cells mostly cause tumor recurrence. In addition, chemotherapy after transurethral resection causes high side effects, and lack of selectivity, and low sensitivity in sensing. Therefore, it is essential to improve new procedures for the diagnosis and treatment of BC. Nanotechnology has recently sparked an interest in a variety of areas, including medicine, chemistry, physics, and biology. Nanoparticles (NP) have been used in tumor therapies as appropriate tools for enhancing drug delivery efficacy and enabling therapeutic performance. It is noteworthy, nanomaterial could be reduced ...
The role of cell adhesion molecules in the progression of bladder urothelial carcinomas
Romanian Journal of Morphology and Embryology
Alteration of the intercellular adhesion system plays an essential role in the initiation and progression of bladder carcinomas. We followed the immunoexpression of adhesion molecules, E-cadherin, β-catenin and Claudin-1, in relation to the histopathological grade and the pT category in a number of 50 urothelial carcinomas of the bladder, based on a final staining score (FSS), calculated on the basis of reaction intensity and labeled cells number. E-cadherin immunoexpression was identified in the membrane of tumor cells, low FSS being associated with invasive high-grade carcinomas. β-catenin reactions were membranous in the case of low-grade noninvasive carcinomas and predominantly cytoplasmic and nuclear in the case of high-grade invasive ones, for which high FSS were associated. Claudin-1 was identified at the membrane level, the high FSS values being more frequent in the case of high-grade invasive carcinomas, although there were no significant statistical associations. Loss of E-cadherin expression and the associated positive linear relation of β-catenin and Claudin-1 indicate the usefulness of the analyzed markers in identifying the invasive aggressive phenotype of urothelial bladder carcinomas.