Development Of Transitional Cell Carcinoma Research Articles

A functional variant rs1464938 in the promoter of fibroblast growth factor 12 is associated with an increased risk of bladder transitional cell carcinoma

Increasing evidence shows that inflammation plays critical roles in the tumorigenesis of bladder cancer. Fibroblast growth factor 12 (FGF12), a kind of inflammatory cytokine, is located in the region of 3q28 that has been demonstrated to be a bladder cancer risk locus by genome wide association study (GWAS). In this study, we aimed to investigate the association of GWAS signal rs710521 and rs884309 and rs1464938 in the promoter of FGF12 with the risk of bladder transitional cell carcinoma (TCC). The polymorphisms were analyzed by using a Taqman assay in 331 TCC patients and 516 age-, gender-, and ethnicity-matched controls. The expression levels of FGF12 mRNA were examined in TCC and non-cancerous normal tissues by using quantitative real-time PCR and the luciferase activity was determined by using the Dual-Luciferase Assay System. The rs1464938 AA genotype and A allele were associated with a significantly increased risk of TCC (AA vs. GG: adjusted OR = 2.54, 95% CI, 1.49–4.35, P < 0.001; AA vs. AG/GG: adjusted OR = 2.25, 95% CI, 1.36–3.71, P = 0.002; A vs. G: adjusted OR = 1.44, 95% CI, 1.15–1.80, P = 0.001, respectively). Haplotype analysis showed that rs884309G- rs1464938A haplotype was associated with an increased risk of TCC (OR = 1.61, 95% CI, 1.23–2.11, P = 0.001). Functional analysis showed that the rs1464938 AG/AA genotypes exhibited higher levels of FGF12 mRNA in TCC tissues and the rs1464938 A allele enhanced FGF12 promoter activity (P < 0.05). These findings suggest that the rs1464938 A allele at the 3q28 locus contribute to the development of TCC by regulating FGF12 expression levels.

Transitional Cell Carcinoma arising in Ovarian Cystic Teratoma: A Rare Case Report

Mature cystic teratoma (MCT) is one of the commonest germ cell neoplasms of the ovary, but only 1–2% gets transformed into a malignant neoplasm. Amongst these, squamous cell carcinoma and adenocarcinoma are most commonly documented in the literature. The development of transitional cell carcinoma is exceptionally uncommon, and only eight cases have been reported in the literature till date as per our knowledge. Here, we report a case of transitional cell carcinoma arising in a dermoid cyst of right ovary in a 48-year-old woman who presented with history of abdominal mass since 3–4 months. Grossly, the ovary was cystic. Microscopically, the cyst wall revealed classic features of MCT and papillary fronds at one place showing features of papillary transitional cell carcinoma (TCC). IHC showed positivity for CK7 and CK20 and negativity for WT-1. When TCC is found as a component of MCT, the origin of the carcinoma should be evaluated.

Gelsolin-like actin-capping protein has prognostic value and promotes tumorigenesis and epithelial-mesenchymal transition via the Hippo signaling pathway in human bladder cancer.

Background:Transitional cell carcinoma (TCC) of the bladder, the major histologic subtype of bladder cancer, is increasing in incidence and mortality, which requires the identification of effective biomarkers. Actin-regulating proteins have recently been proposed as important antitumor druggable targets. As a gelsolin-family actin-modulating protein, CAPG (gelsolin-like actin-capping protein) generated great interest due to its crucial effects in various biological and physiological processes; however, the role and mechanism of CAPG in TCCs remain unknown.Materials and methods:Bioinformatic analysis and immunohistochemistry of clinical specimens were performed to detect the expression level of CAPG. Both in vitro and in vivo assays were used to determine the oncogenic effect of CAPG in TCCs. Male 4–5-week-old BALB/c nude mice were used for in vivo tumorigenesis assays, while SCID mice were used for in vivo metastatic assays. Affymetrix microarray was used to identify the underlying molecular mechanism. Western blot and immunofluorescence were used to validate the expression and localization of proteins.Results:CAPG was frequently upregulated in TCCs and associated with clinical aggressiveness and worse prognosis. Functional assays demonstrated that CAPG could contribute to the tumorigenesis, metastasis and epithelial-mesenchymal transition (EMT) of TCCs both in vitro and in vivo. A novel mechanism that CAPG promoted TCC development via inactivating the Hippo pathway, leading to a nucleus translocation of Yes-associated protein was suggested.Conclusions:The current study identified CAPG as a novel and critical oncogene in TCCs, supporting the pursuit of CAPG as a potential target for TCC intervention.

Upregulated glutathione transferase omega-1 correlates with progression of urinary bladder carcinoma

ABSTRACTObjectives: Newly discovered glutathione transferase omega 1 (GSTO1-1) plays an important role in the glutathionylation cycle, a significant mechanism of protein function regulation. GSTO1-1 expression pattern has not been studied in transitional cell carcinoma (TCC), as yet.Methods: A total of 56 TCC tumor and corresponding non-tumor specimens were investigated. Glutathione content and thioltransferase activity were measured spectrophotometrically. Protein-glutathione mixed disulfides were measured fluorimetrically. GSTO1-1 expression was determined by immunoblot and qPCR. Immunoprecipitation with GSTO1-1 antibody was followed by immunoblot using anti-GSTO1, GSTP1, c-Jun, JNK, Akt, phospho-Akt, and ASK1 antibody, while for the total S-glutathionylation levels non-reducing electrophoresis was performed.Results: The contents of reduced glutathione and thioltransferase activity were significantly increased in tumor compared to non-tumor tissue. The increased GSTO1 expression in tumor tissue showed clear correlation with grade and stage. However, decreased total protein glutathionylation level in tumor compared to non-tumor samples was found. Immunoprecipitation has shown an association of GSTO1-1 with GSTP1, Akt, phospho-Akt, and ASK1 proteins.Conclusions: GSTO1 deglutathionylase activity suggests its potential important role in redox perturbations present in TCC. Increased GSTO1-1 expression might contribute to TCC development and/or progression supporting the notion that GSTO1-1 may be a promising novel cancer target.

Peritoneal carcinomatosis in urinary bladder cancer: A case report

Transitional cell carcinoma (TCC) is associated with malignant tumors that arise from stem cells near the basement membrane of the epithelial bladder surface. Exposure of carcinogens to the urothelium leads to genetic alterations in cells, increasing the malignant potential. Of the different environmental exposures, cigarette smoke is considered the greatest risk factor in the development of TCC in the United States. Pathological tumor growth patterns are classified as papillary (most common), sessile/mixed, or nodular, with TCC often arising via the flat or papillary pathways. Most commonly, TCC involves the papillary pathway, with formation of a tumor projecting into the bladder lumen and possible invasion of the lamina propria and bladder muscle. Constitutional symptoms such as weight loss, fatigue, along with lower abdominal pain or bone pain are often indicative of metastatic disease and poor prognosis. Metastasis of TCC most commonly occurs to the lungs, liver, or bone. Metastasis to the peritoneum with abdominal ascites, however, is very rare. In our case, TCC with peritoneal carcinomatosis and ascites in a 48-year-old male smoker can be appreciated. Abdominal pain with nausea, weight loss, and hematuria lead to computed tomography (CT) scan, which depicted ascites, omental thickening, and soft tissue mass in the urinary bladder. Biopsy and pathologic cytology revealed high-grade TCC with papillary morphology. Transitional cell carcinoma with peritoneal carcinomatosis is a rare cause of bladder carcinoma. Poor prognosis is due to advanced disease stage, poor performance status, and metastasis. Effective treatment often cannot be initiated, because of the grim presentation. (This page in not part of the published article.) International Journal of Case Reports and Images, Vol. 7 No. 11, November 2016. ISSN – [0976-3198] Int J Case Rep Images 2016;7(11):749–753. www.ijcasereportsandimages.com Khosla et al. 749 CASE REPORT OPEN ACCESS Peritoneal carcinomatosis in urinary bladder cancer: A case report Manraj Khosla, Ali Imran, Panagiotis Fidias

GSTO1*C/GSTO2*G haplotype is associated with risk of transitional cell carcinoma of urinary bladder.

To clarify the role of genetic polymorphisms of GSTO1 (rs4925) and GSTO2 (rs156697) in individual susceptibility to urinary bladder cancer. Case-control study consisting of 187 patients with histologically confirmed transitional cell carcinoma (TCC) of urinary bladder and 140 age- and gender-matched cancer-free controls was carried out. Genotyping of GSTO1 and GSTO2 was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). We found that carriers of mutant GSTO2*G/G genotype were at increased risk of the development of TCC (OR 2.6, 95% CI 1.2-5.8, p = 0.041), while GSTO1 rs4925 polymorphism was not significantly associated with TCC risk (p = 0.450). According to smoking status, smokers with GSTO2*G/G genotype had significantly higher risk of TCC of urinary bladder (OR 4.3, 95% CI 1.6-11.2, p = 0.003) compared to wild-type carriers with no smoking history. We further analyzed the effects of GSTO1/GSTO2 haplotypes on TCC risk, based on the linkage disequilibrium found for GSTO1 (rs4925) and GSTO2 (rs156697) (D' = 0.309, p = 0.001). The study subjects with GSTO1*C/GSTO2*G (GSTO1 wild-type/GSTO2 mutant) haplotype were at the highest risk of the development of transitional cell carcinoma of urinary bladder (OR 2.8, 95% CI 1.5-5.2, p = 0.002). Our results indicate that GSTO1*C/GSTO2*G haplotype is associated with increased risk of TCC. The modifying effect of GSTO2*G/G genotype on individual susceptibility to TCC is more pronounced, when associated with smoking.

Mature cystic teratoma with malignant transformation of teratomatous urothelial cells: Rare case presentation

The occurrence of malignancies in somatic elements of mature cystic teratoma of ovary is rare. The malignancies that may be encountered in dermoid cyst include squamous cell carcinoma, adenocarcinoma, adenosquamous carcinoma, melanoma, sarcoma, carcinoid, and germ cell neoplasms. The development of transitional cell carcinoma (TCC) in dermoid cyst is extremely rare with only four such cases having been reported in literature so far. Here we report the fifth case of such an occurrence in a 50-year-old postmenopausal multiparous female patient. She presented with pain and gradual swelling of abdomen for 1 month. Abdominal computed tomography revealed a solid space occupying lesion with few cystic components at right pelvis, raising the possibility of an ovarian neoplasm. The level of CA-125 was slightly raised (56∙45 U/ml). Total abdominal hysterectomy and bilateral salpingo-oopherectomy was performed. Microscopic examination showed cyst wall lined by stratified squamous epithelium. Beneath the cyst wall, a tumor mass was present, histological features of which resembled that of high-grade TCC (stage pT1aNXMX). On immunohistochemical analysis, the tumor was found to be positive for CK7 and CK20 and negative for WT-1. These results were consistent with a diagnosis of TCC arising in urothelium of mature cystic teratoma. Reporting of such extremely rare cases is important for the assessment of prognostic factors and treatment protocols.

Unexpected events of ureteric schistosomiasis in Saudi Arabia

The incidence of schistosomiasis in the Kingdom of Saudi Arabia (KSA) is low; although the presence of ureteric schistosomiasis is low, its presence in association with the other complications is considered a rare event among males, especially in a country like KSA. The objective was to study a 56-year-old Saudi Arabian male, who presented with scistosomiasis in ureter with the development of transitional-cell carcinoma instead of squamous-cell carcinoma. An investigation was carried out in this case by repeated cystoscopes and biopsies for histopathological examinations, as well as several imaging procedures such as plain X-ray, CT, and MRI along with several lab assessments. The presence of all these complications in this patient makes this a case of special interest. This will reflect on conducting further research workup for accurate estimation of schistosomiasis in Saudi Arabia.

Inhibition of Mouse Urinary Bladder Carcinogenesis by Açai Fruit (Euterpe oleraceae Martius) Intake

Açai, fruit from Euterpe oleraceae Martius, is consumed in natura and in a variety of beverages and food preparations and possesses several potential antioxidant compounds. In a first study for anticarcinogenicity screening, male Swiss mice (n = 20/per group) were chemically-induced to urothelial bladder carcinogenesis for 10weeks and received a standard diet or a standard diet containing 2.5 and 5% spray-dried açai pulp (AP) for 10weeks. At week 20, the incidence of simple and nodular hyperplasia and the incidence and multiplicity of transitional cell carcinoma (TCC) were evaluated. In a second study for antigenotoxicity screening, male Swiss mice (n = 6/per group) were fed standard diet or standard diet containing 5% AP for threeweeks. Urothelial cell suspensions were obtained and challenged with H(2)O(2) for induction of DNA damage and analyzed by comet assay. Overall, dietary 5% AP reduced TCC incidence and multiplicity (p = 0.019 and p = 0.015, respectively) and tumor cell proliferation and p63 expression (p = 0.02 and p = 0.007, respectively), Furthermore, the group fed the 5% AP presented a significant reduction (p < 0.01) in DNA damage induced by H(2)O(2), a notable oxidant agent. The results suggest that the spray-dried açai pulp used here inhibits the TCC development in male Swiss mice, probably due to its potential antioxidant action.

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Aim The clinical behavior of the human urinary bladder papillary transitional cell carcinoma (TCC) probably results from a complex interaction between carcinogenic insults and host resistance during the patient’s life. While the main recognized risk factor is smoking, little information exists about the susceptibility to TCC development. The human leukocyte antigen E (HLA-E) molecule plays an important role both as a modulator of natural killer (NK) cell activity by interacting with CD94-NKG2A receptor in the innate immunity pathway and as an antigen-presenting molecule triggering a specific immune response, thus it may influence tumor immune surveillance. The aim of this study was to explore a possible influence of five HLA-E variation sites, one in exon 2 at position 424 (rs1059510), three at exon 3 in the positions 756 (rs1264457), 887 (rs41560815) and 906 (rs41562314) and one in exon 4 at the position 1691 (rs17875370), on the susceptibility to urinary bladder TCC development and progression in Brazilian subjects. Methods These variation sites were evaluated by means of direct sequencing of PCR products using an ABI3100 Genetic Analyzer. On this bases, 105 healthy donors and 73 patients with historical of TCC, both from Ribeirao Preto City, Southeast Brazil, were evaluated for the variation sites described above. Results Only two of the five variation sites evaluated were polymorphic in an admixed population such as Brazilians, i.e., positions +424 and +756. Considering these, the allele and genotype distributions in patients and controls were very similar and no significant difference was detected, even when patients were compared as a whole or stratified into high and low grade tumor patients. Conclusions In conclusion, although previous associations have been described evolving neighboring genes such as HLA-A and HLA-G, the HLA-G locus seems to be not associated with TCC susceptibility and/or progression to aggressive tumors.

Antioncogenic Effects of Transient Receptor Potential Vanilloid 1 in the Progression of Transitional Urothelial Cancer of Human Bladder

The progression of normal cells to a tumorigenic and metastatic state involves the accumulation of mutations in multiple key signaling proteins, encoded by oncogenes and tumor suppressor genes. Recently, members of the TRP channel family have been included in the oncogenic and tumor suppressor protein family. TRPM1, TRPM8, and TRPV6 are considered to be tumor suppressors and oncogenes in localized melanoma and prostate cancer, respectively. Herein, we focus our attention on the antioncogenic properties of TRPV1. Changes in TRPV1 expression occur during the development of transitional cell carcinoma (TCC) of human bladder. A progressive decrease in TRPV1 expression as the TCC stage increases triggers the development of a more aggressive gene phenotype and invasiveness. Finally, downregulation of TRPV1 represents a negative prognostic factor in TCC patients. The knowledge of the mechanism controlling TRPV1 expression might improve the diagnosis and new therapeutic strategies in bladder cancer.

The management of transitional cell carcinoma (TCC) in a European regional renal transplant population

In the West, transitional cell carcinoma (TCC) in renal transplant patients is uncommon, but aggressive. Conversely, it appears to be frequent in the Far East, necessitating aggressive surgical approaches such as prophylactic nephroureterectomy. There are few European case series to date. TCC in the present population was predominantly low-grade and superficial, with no progression in patients with those tumours. Endoscopic management was sufficient for most patients. The behaviour of TCC in the present population was much less aggressive than that described in the Far East. Altering immunosuppression regimes may have a role to play in managing bladder cancer in renal transplant patients. To examine the clinical characteristics, management and long-term outcomes of patients with transitional cell carcinoma (TCC) who also have had renal transplantation. A retrospective case note review was performed for the 15-year period 1995-2009. Searches from three different urological centres in the UK, using multiple sources, yielded 1647 patients with renal transplants, 12 of whom had TCC. Eight cases were identified who developed de novo TCC after transplantation (0.48%). Four patients had pre-existing TCC who then had renal transplantation. The current literature was reviewed. In the eight de novo TCC cases, the bladder was the site in all with no upper tract TCC; seven were superficial (pTa/T1) and five were low grade (G1/2). The mean time to development of TCC after transplant was 5 years, with a mean follow-up of 11 years. There was no progression in low-grade superficial disease that was managed endoscopically. The 5- and 10-year overall survival was 83% and 72%, respectively. In patients with pre-existing TCC prophylactic bilateral nephroureterectomy before transplantation was performed once. There was progression of superficial disease whilst on immunosuppression in one patient. Sirolimus was used in patients with TCC and reports suggest this may have a role to play in modifying malignancy in this setting. The number of patients involved in studies particularly focusing on TCC in renal transplantation is small (136 patients), with 60% from China/Taiwan where there is a high incidence of upper tract TCC and high-grade muscle-invasive disease. Although this is one of the largest European case series of renal transplant patients with TCC, the numbers are small making clear conclusions difficult. The frequency of TCC in our renal transplant population is low, consistent with previous studies. However, contrary to prior studies, TCC after renal transplantation in this European population was predominantly superficial, low-grade, non-progressive and confined to the bladder. Altering immunosuppression regimes in patients with TCC may have a role to play, although further work is required to clarify and substantiate this.

Transitional Cell Carcinoma of the Native Urinary Tract After Kidney Transplantation: Recommendations Following a Long-Term Retrospective Analysis

IntroductionThe aim of the current study was to explore the clinico-oncological characteristics, and the therapeutic and survival parameters, of renal transplant recipients who developed de novo transitional cell carcinoma (TCC) over a 30-year period at the authors’ center. MethodsRetrospective analysis of records from all registered patients who underwent kidney transplantation at the center between November 1979 and January 2010 who developed de novo TCC of the urinary tract. ResultsFrom all 2001 patients analyzed during the study period, 21 recipients developed 19 TCCs of the bladder and 6 TCCs of the upper urinary tract. Among bladder TCCs, 13 (68.4%) cases were confined to mucosa (pTa or carcinoma in situ, n=7) or submucosa (pT1, n=6) and 6 others (31.6%) infiltrated the detrusor muscle (≥p T2); the grading distribution was 5 cases of G1, 6 of G2 and 8 of G3. All recurrent cases (n=8) revealed local or systemic progression. The overall and tumor-specific patient survival rates were 80.2%, 54.0% and 30.0% and 84.9%, 67.4% and 58.9% for 1, 5 and 10 years, respectively. ConclusionsIn the light of the observed increased aggressiveness of TCC in renal transplant recipients, more frequent examinations and possibly more invasive follow-up protocols should be considered for patients with 1 or more risk factors for development of TCC. Urine cytology (including ureteral wash cytology) may be used as a reliable diagnostic tool in these patients. Prophylactic contralateral nephroureterectomy might be an option in patients at high risk.

Phase II trial of everolimus monotherapy in the palliative treatment of patients with metastatic transitional cell carcinoma (TCC) after failure of platinum-based therapy: Activity and biomarkers.

4622 Background: Dysregulation of mammalian Target of Rapamycin (mTOR) pathway plays a role in TCC development. We assessed the efficacy and toxicity of Everolimus (E), an oral mTOR inhibitor, in patients with palliative TCC in a phase II study and explored potential predictive plasma biomarkers of E activity. Methods: Patients (pts) with metastatic TCC after failure of platinum chemotherapy received E 10 mg/d continuously until progressive disease (PD) or unacceptable toxicity. The primary end point was stable disease (SD) or partial response (PR) at 8 wks according to RECIST criteria (centrally reviewed). A 2-stage design was used (Simon; a=0.15, b=0.1, P0=10%, P1=25%). Plasma samples were collected on day 1 (baseline before treatment), day 28 (during treatment), and at PD. Different angiogenesis-related proteins were analyzed in plasma by cytokine arrays and ELISA. Results: 37 pts were included. 8 pts were not evaluable by imaging at 8 wks due to rapid clinical deterioration (3), death (4), toxicity (1) and were considered in PD. 1 pt was lost at follow-up. Confirmed PR was observed in 2 pts, SD in 8, and PD in 27, resulting in a disease control rate of 27% at 8 wks. In 3 out of these 8 pts with SD, a decrease in the sum of the largest diameters of the target lesions was recorded (1%, 2%, 9%). SD was maintained for 16 weeks in 4 pts, for 24 weeks in 2, and for 52 weeks in 1. Toxicities of E were as reported previously. Plasma analysis showed that E induced changes in angiostatin (+ 114%; p<0.0001) and in PDGF-AB (- 68%; p=0.007) levels. High baseline angiopoietin-1 (Ang-1) levels were correlated to PR/SD (p=0.0057). Pts in SD/PR showed an Ang-1 decrease (- 64%; p=0.02) on day 28 compared to pts in PD (+ 26%; p=0.49). Furthermore, PD was associated with an endoglin increase on day 28 (+ 63%; p=0.0023). Interestingly, in the 2 pts with a PR, an increase in Ang-1, PDGF-AA and PDGF-AB levels was observed at the time of PD compared to those previously detected on day 28. Conclusions: E monotherapy showed moderate activity in advanced TCC. Ang-1, PDGF-AB and endoglin are potential biomarkers that should be investigated further in larger clinical trials.

Identification of genes correlated with early-stage bladder cancer progression.

Transitional cell carcinoma (TCC) of the bladder ranks fourth in incidence of all cancers in the developed world, yet the mechanisms of its origin and progression remain poorly understood. There are also few useful diagnostic or prognostic biomarkers for this disease. We have combined a transgenic mouse model for invasive bladder cancer (UPII-SV40Tag mice) with DNA microarray technology to determine molecular mechanisms involved in early TCC development and to identify new biomarkers for detection, diagnosis, and prognosis of TCC. We have identified genes that are differentially expressed between the bladders of UPII-SV40Tag mice and their age-matched wild-type littermates at 3, 6, 20, and 30 weeks of age. These are ages that correspond to premalignant, carcinoma in situ, and early-stage and later stage invasive TCC, respectively. Our preliminary analysis of the microarray data sets has revealed approximately 1,900 unique genes differentially expressed (> or =3-fold difference at one or more time points) between wild-type and UPII-SV40Tag urothelium during the time course of tumor development. Among these, there were a high proportion of cell cycle regulatory genes and a proliferation signaling genes that are more strongly expressed in the UPII-SV40Tag bladder urothelium. We show that several of the genes upregulated in UPII-SV40Tag urothelium, including RacGAP1, PCNA, and Hmmr, are expressed at high levels in superficial bladder TCC patient samples. These findings provide insight into the earliest events in the development of bladder TCC as well as identify several promising early-stage biomarkers.

SOX2 modulates alternative splicing in transitional cell carcinoma

Aberrant alternative splicing of key cellular regulators may play a pivotal role in cancer development. To investigate the potential influence of altered alternative splicing on the development of transitional cell carcinoma (TCC), splicing activity in the TCC cell lines TSGH8301 and BFTC905 was examined using the SV40-immortalized uroepithelial cell line SV-HUC-1 as a reference. Our results indicate a significant alteration in splice site selection in the TCC cell lines. By gene expression profiling and subsequent validation, we discovered that sex-determining region Y-box protein 2 (SOX2) is specifically upregulated in BFTC905. Furthermore, ectopic expression of SOX2 modulates alternative splicing of the splicing reporter in vivo. More significantly, using an in vitro pull-down assay, it was found that SOX2 exhibits RNA-binding capability. Our observations suggest that SOX2 modulates alternative splicing by functioning as a splicing factor.

Human papillomavirus is detected in transitional cell carcinoma arising in renal transplant recipients

We investigated the role of human papillomavirus (HPV) in the development of transitional cell carcinoma (TCC) arising in renal transplant recipients. Genomic DNA was extracted from 10 microm paraffin embedded sections of five TCCs arising in five renal transplant recipients using the QIAamp DNA mini kit according to the manufacturer's instructions. beta-globin PCR was performed to test DNA adequacy. Samples were tested for the presence of HPV DNA by broad spectrum HPV PCR method using non-biotinylated SPF10 primers (SPF1A, SPF1B, SPF1C, SPF1D, SPF2B, SPF2D) which amplify a short 65 bp fragment. Positive bands were identified on a 3% gel. Positive samples underwent a second HPV PCR and were amplified using biotinylated SPF10 primer set, which amplifies the same 65 bp region of the L1 open reading frame. INNO-LiPA line probe assay was then performed to genotype the samples which uses a reverse hybridisation principle. Four of five TCCs examined were positive for HPV. The high risk HPV16 was detected in three cases whereas in the fourth case an unclassifiable HPV genotype was present. In all DNA samples, beta-globin amplification was successful. Our results indicate that HPV and in particular HPV16 may play an aetiological role in the development of TCC in renal transplant patients.

Mutational analysis of the BRAF gene in transitional cell carcinoma of the bladder

Mutational activation of the MAP kinase pathway is frequently found in many types of cancer. Recently, activating mutations in the BRAF gene, an important activator of this pathway, have been described in several tumor types including melanoma, colorectal and papillary thyroid cancer. The most frequent mutation in exon 15 (V600E) as well as several other mutations within exons 11 and 15 result in constitutive activation of the oncoprotein. Our study aimed to investigate BRAF mutations in 30 human bladder tumors and their adjacent normal tissues. The V600E mutation was screened by PCR/RFLP and exons 11, 14 and 15 of BRAF including intron-exon boundaries were sequenced. We detected two tumor specimens bearing two different mutations, both of which were found in exon 15. One sample showed the T1799A (V600E) and the other the G1798T (V600L) mutation. The first specimen was stage pT1a and grade II, whereas the second was stage pT2b and grade III. No mutations within the coding region of exons 11, 14, 15 and the intron-exon junctions for the remaining samples were found. Our results suggest that involvement of BRAF mutations in the development of transitional cell carcinoma of the bladder is infrequent.

Can We Use Single Nucleotide Polymorphism and Runt Domain Transcription Factor 3 Methylation as Tumor Markers for Bladder Cancer?

Purpose: Although many tumor markers have been evaluated in relation to bladder cancer, none of these biomarkers reported to date has shown sufficient sensitivity and specificity for the detection of the whole spectrum of bladder cancer diseases in routine clinical practice. The limited value of the established prognostic markers requires analysis of new molecular parameters of interest for predicting the prognosis of bladder cancer patients. Materials and Methods: We conducted a review of the literature with a focus on recent advances in genetic polymorphism and hypermethylation events in relation to bladder transitional cell carcinoma. Results: Recently, there has been major progress in both genetic polymorphism in relation to bladder cancer and molecular genetic and epigenetic changes leading to the development of transitional cell carcinoma. However, studies on numerous single-nucleotide polymorphisms in relation to bladder cancer have provided only a few genetic polymorphisms with only marginal information on patients’ prognosis. For this reason, interest is increasing in epigenetic changes in bladder cancer. The epigenetic silencing of tumor suppressor genes is interesting from a clinical standpoint because of the possibility to reverse the epigenetic changes and thus restore gene function to a cell. Treatment with DNA methylation inhibitors can restore the activities of dormant genes and decrease the growth rate of cancer cells in a heritable fashion. Conclusions: Epigenetic modification may be possible to partially reverse the cancer phenotype, and this will eventually lead to targeted therapy tailored toward specific molecular therapy in the near future. (Korean J

Mutational Analysis of the BRAF Gene in Transitional Cell Carcinoma of the Bladder

Mutational activation of the MAP kinase pathway is frequently found in many types of cancer. Recently, activating mutations in the BRAF gene, an important activator of this pathway, have been described in several tumor types including melanoma, colorectal and papillary thyroid cancer. The most frequent mutation in exon 15 (V600E) as well as several other mutations within exons 11 and 15 result in constitutive activation of the oncoprotein. Our study aimed to investigate BRAF mutations in 30 human bladder tumors and their adjacent normal tissues. The V600E mutation was screened by PCR/RFLP and exons 11, 14 and 15 of BRAF including intron-exon boundaries were sequenced. We detected two tumor specimens bearing two different mutations, both of which were found in exon 15. One sample showed the T1799A (V600E) and the other the G1798T (V600L) mutation. The first specimen was stage pT1a and grade II, whereas the second was stage pT2b and grade III. No mutations within the coding region of exons 11, 14, 15 and the intron-exon junctions for the remaining samples were found. Our results suggest that involvement of BRAF mutations in the development of transitional cell carcinoma of the bladder is infrequent.