Expression In Transitional Cell Carcinoma Research Articles

600 - Prostate specific membrane antigen (PSMA) expression in transitional cell carcinoma (TCC) – who puts the S in PSMA?

600 - Prostate specific membrane antigen (PSMA) expression in transitional cell carcinoma (TCC) – who puts the S in PSMA?

CD10 and CA19.9 immunohistochemical expression in transitional cell carcinoma of the urinary bladder

Background: Transitional cell carcinoma of the bladder is the most common malignancy affecting the urinary tract ranking the 5th among males and the 9th among females’ cancers in Iraq. The prognosis depends largely on the histological grade and stage of the tumor at diagnosis; however, there is no reliable parameter predicting the risk of recurrence or progression; molecular and immunological markers may be required to estimate the individual prognosis of patients as well as for effective diagnosis and treatment.Objectives: To evaluate CD10 and CA19.9 immunohistochemical expression in transitional cell carcinoma of the urinary bladder and to correlate this expression with the grade and stage of the tumor.Materials and Methods: This study was retrospectively designed. Forty-nine cystoscopy specimens of urothelial carcinoma of the bladder were retrieved from the archival materials of the Specialized Surgical Hospital and Al-Khadhmiya Teaching Hospital in Baghdad for the period from January 2010 to June 2011. Three sections of 5-μm thickness were taken from each case. One section was stained with Hematoxylin and Eosin; the other two were stained immunohistochemically with CA19.9 and CD10.Results: Immunohistochemical expression of CA19.9 and CD10 had a significant correlation with WHO 2004 grade of urothelial carcinoma. There was no significant correlation between CA19.9 and CD10 immunohistochemical expression with stage.Conclusions: CA19.9 and CD10 immunohistochemical expression could be of value in assisting the differentiation between high and low-grade urothelial carcinoma cases and consequently in determining the prognosis in such cases.

A study of p53 expression in transitional cell carcinoma of urinary bladder in Erbil

Background and objectives: This study aimed to evaluate p53 protein expression in both normal bladder epithelium and in cases of transitional cell carcinoma (TCC) of urinary bladder by immunohistochemical study and to correlate p53 expression in urothelial cancer with other clinico-pathological parameters. Methods:This is a retrospective and prospective study for sample collection during the period from January 2006-May 2009. The samples studied included 105 formalin fixed, paraffin embedded urinary bladder tissue specimens; they consisted of the following diagnostic categories: chronic non specific cystitis (n=5) and urothelial cancer (n=100). In this study the nuclear p53 protein expression was detected in tissue samples by Dako Cytomation. LSAB + System-HRP staining protocol using monoclonal mouse anti human protein DO-7.  Results:None of the chronic non specific cystitis cases showed p53 nuclear immunostaining, while 93% of urothelial cancer specimens examined showed immunopositivity for p53 protein. In this study, a statistically significant correlation was observed between p53 overexpression rate with the tumor grade (p= <0.001) and histological architecture (p= 0.023), but not with other clinico-pathological parameters like age and gender. Conclusion: Results of the present study showed the validity and simplicity of application of immunohistochemistry in determining the status of p53 protein expression. The results suggest that p53 overexpression is strongly associated with the aggressiveness of urothelial cancer.

Epigenetic regulation of vascular endothelial growth factor a dynamic expression in transitional cell carcinoma

Vascular endothelial growth factor A (VEGF-A) is a key mediator in the neovascularization of cancers. We have found that VEGF-A was expressed at significantly higher levels in high-grade transitional cell carcinoma (TCC) cells than low-grade TCC cells in our previous study. In the present study, promoter methylation pattern was assessed and quantified by bisulfite genomic sequencing (BGS) and specific VEGF-A CpG sites in low-grade, but not in high-grade, TCC cells were observed. Reporter assays indicated that hypermethylation of nine CpG sites can inhibit the transcriptional activity of the VEGF-A gene. Subsequent chromatin immunoprecipitation (ChIP) assay revealed down-regulation of transcription activity of VEGF-A with increasing binding of methyl-CpG-binding protein 2 (MBD2) and trimethyl-histone H3 (Lys9) proteins to these CpG sites in low-grade TCC cells during hypermethylation. Furthermore, treatment of low-grade TCC cells with DNA methyltransferase inhibitor and histone deacetylase inhibitor can restore the expression of VEGF-A and promote the invasive ability of low-grade TCC cells. Hypermethylation with lower expression levels of VEGF-A in low-grade TCC tumors than high-grade TCC tumors was also confirmed in clinical specimens by reverse transcriptase-PCR and pyrosequencing analyses. Our findings are the first results indicating that VEGF-A expression is suppressed in low-grade TCC tumors by promoter hypermethylation. This offers a new perspective on the role of VEGF-A in TCC tumor behavior.

Differential Protein Expression Profiling by iTRAQ-Two-dimensional LC-MS/MS of Human Bladder Cancer EJ138 Cells Transfected with the Metastasis Suppressor KiSS-1 Gene

KiSS-1 is a metastasis suppressor gene reported to be involved in the progression of several solid neoplasias. The loss of KiSS-1 gene expression has been shown to be inversely correlated with increasing tumor stage, distant metastases, and poor overall survival in bladder tumors. To identify the molecular pathways associated with the metastasis suppressor role of KiSS-1 in bladder cancer, we carried out a proteomics analysis of bladder cancer cells (EJ138) transiently transfected with a vector encompassing the full-length KiSS-1 gene using an iTRAQ (isobaric tags for relative and absolute quantitation) approach. Protein extracts collected after 24- and 48-h transfection were fractionated and cleaved with trypsin, and the resulting peptides were labeled with iTRAQ reagents. The labeled peptides were separated by strong cation exchange and reversed phase LC and analyzed by MALDI-TOF/TOF MS. Three software packages were utilized for data analysis: ProteinPilot for identification and quantification of differentially expressed proteins, Protein Center for gene ontology analysis, and Ingenuity Pathways Analysis to provide insight into biological networks. Comparative analysis among transfected, mock, and empty vector-exposed cells identified 1529 proteins with high confidence (>99%) showing high correlation rates among replicates (70%). The involvement of the identified proteins in biological networks served to characterize molecular pathways associated with KiSS-1 expression and to select critical candidates for verification analyses by Western blot using independent transfected replicates. As part of complementary clinical validation strategies, immunohistochemical analyses of proteins regulated by KiSS-1, such as Filamin A, were performed on bladder tumors spotted onto tissue microarrays (n = 280). In summary, our study not only served to uncover molecular mechanisms associated with the metastasis suppressor role of KiSS-1 in bladder cancer but also to reveal the biomarker role of Filamin A in bladder cancer progression and clinical outcome.

Exon 2 Methylation Inhibits hepaCAM Expression in Transitional Cell Carcinoma of the Bladder

Aim: We aimed to investigate the mechanisms of hepaCAM inactivation in transitional cell carcinoma of the bladder through the analysis of hepaCAM exon 2 methylation. Methods: The methylation of hepaCAM exon 2 and the expression of hepaCAM were determined by methylation-specific restriction PCR assay and RT-PCR in bladder cancer cells (T24, BIU-87) as well as in 55 paired bladder cancer specimens. The methylated bladder cancer cells were treated with 5-Aza- 2′-deoxycytidine (5-Aza-CdR), a demethylating agent. MTT was used to detect the proliferation of T24 and BIU-87 cells. Results: The proliferation of T24 and BIU-87 cells was suppressed by treatment with different concentrations of 5-Aza-CdR; the expression of hepaCAM was absent in T24 and BIU-87 cells, and we found that exon 2 of hepaCAM was methylated in the 2 cells. hepaCAM mRNA was re-expressed and the methylation status of hepaCAM exon 2 was reversed after treatment with 5-Aza-CdR. The expression of hepaCAM mRNA in bladder cancer tissues was significantly lower than that in adjacent tissues. The methylation rate of hepaCAM exon 2 was significantly higher in bladder cancer tissues than in adjacent tissues. The methylation of hepaCAM exon 2 was related to hepaCAM expression in bladder cancer tissues. Conclusions: Downregulation of hepaCAM expression plays an important role in the tumorigenesis and development of bladder cancer. DNA methylation may be important for downregulation of hepaCAM expression in bladder cancer.

99 Cycloxygenase-2 (COX2) expression in transitional cell carcinoma of the bladder does not confer independent prognostic properties

99 Cycloxygenase-2 (COX2) expression in transitional cell carcinoma of the bladder does not confer independent prognostic properties

Relationship Between Cysteinyl-Leukotriene-1 Receptor and Human Transitional Cell Carcinoma in Bladder

To investigate the leukotriene (LT) D(4) (LTD(4)) receptor (cysteinyl-LT(1) receptor [CysLT(1)R]) expression in transitional cell carcinoma (TCC) of the bladder, as well as the effects of the CysLT(1)R antagonist on cell proliferation in TCC cell lines. The metabolism of arachidonic acid by either cyclooxygenase or lipoxygenase is thought to play an important role in carcinogenesis. LTD(4) is a pro-inflammatory mediator derived from arachidonic acid through various enzymatic steps, and 5-lipoxygenase is an important factor in generating LTD(4). CysLT(1)R expression in TCC tissue and normal bladder tissue was examined. CysLT(1)R expression was detected using immunohistochemistry. The effects of the CysLT(1)R antagonist on TCC cell growth were examined by 3-(4,5-dimethylthiazol-2-thiazolyl)-2,5-diphenyltetrazolium bromide assay and reverse transcriptase-polymerase chain reaction. Flow cytometry was used to determine whether the CysLT(1)R antagonist induced apoptosis. Initially, only slight CysLT(1)R expression was detected in normal bladder tissues and marked CysLT(1)R expression was detected in the TCC tissues. CysLT(1)R expression was greater in high-grade cancer than in low-grade cancer. Furthermore, CysLT(1)R expression was also greater in advanced-stage cancer than in early-stage cancer. Finally, the CysLT(1)R antagonist caused marked inhibition of TCC cells by inducing early apoptosis. CysLT(1)R was induced in TCC. The results suggest that the CysLT(1)R antagonist might mediate potent antiproliferative effects on TCC cells. Thus, the target of the CysLT(1)R is potentially a new therapy in the treatment of TCC.

Enhanced GSTP1 expression in transitional cell carcinoma of urinary bladder is associated with altered apoptotic pathways

Objectives Glutathione S-transferase P1 (GSTP1) provides an important link between activity of regulatory stress kinases and apoptotic pathways. It can be hypothesized that up-regulated GSTP1, in TCC, might enhance apoptosis inhibition. We aimed to establish whether relationship between GSTP1 expression and executive (pro-caspase 3, cleaved caspase 3) and regulatory (Bcl-2) apoptotic pathways in TCC exists. Materials and Methods Samples were obtained from 84 TCC patients (41 consecutive patient with muscle noninvasive and 43 consecutive patients with muscle invasive TCC tumors), who underwent surgery at the Institute of Urology and Nephrology, Clinical Centre of Serbia, during 2006 and 2007. Expression of GSTP1, pro-caspase 3 (CPP32), and Bcl-2, as well as cleaved caspase-3 labeling index (LI) were determined by immunocytochemistry. Levels of expression were correlated with tumor stage, grade, and invasiveness. Results GSTP1 protein expression was demonstrated in all tumor samples examined. According to GSTP1 status, all tumors were divided into groups with low, moderate, or high GSTP1 status. Expression of CPP32 and cleaved caspase 3 was positive in 80% of TCC patients. Their levels differed significantly between groups with various GSTP1 expression ( P < 0.05), with the lowest CPP32 expression and cleaved caspase 3 LI in tumors with high GSTP1 status. Moreover, significant negative correlation was found between GSTP1 level and cleaved caspase 3 LI (r = –0.459, P = 0.041). The positive rate of Bcl-2 protein expression was 48%. Most of the Bcl-2 positive patients exhibited at the same time high GSTP1 positivity ( P = 0.078). Significant association with tumor grade and stage was found for all examined parameters except for CPP32 regarding tumor grade. Conclusions Based on results obtained, we conclude that enhanced GSTP1 expression in TCC of urinary bladder is associated with altered apoptotic pathways. Molecular interplay between GSTP1 and members of apoptotic cascade might, at least partially, play a role in development of invasive characteristics of TCC.

Loss Expression of Uroplakin III is Associated with Clinicopathologic Features of Aggressive Bladder Cancer

Tissue-specific expression is of key importance in gene therapy and can be achieved by using tissue-specific promoters to drive therapeutic gene expression. The uroplakin (UP) promoter is a powerful tool for bladder cancer gene therapy, but the role of UP protein in the bladder remains unknown. This study aimed to detect UP III expression in transitional cell carcinoma (TCC) of the bladder and to determine whether the role of UP III heterogeneity is associated with predicting disease recurrence and patient survival. Immunohistochemical staining for UP III was carried out in 92 archival radical cystectomy and 38 normal specimens and correlated with pathologic features and clinical outcomes. UP III expression was significantly decreased in bladder cancer tissues compared with normal controls (P <0.0001). Loss of UP III expression was associated with high-grade, muscle-invasive cancer, lymphovascular invasion (P <0.01), and decreased cancer-specific survival at a median follow-up of 25.3 months (P = 0.04). When adjusted for the effects of standard pathologic features, only lymph node metastases were associated with bladder cancer progression (P = 0.01) and mortality (P = 0.04). Loss of UP III expression is associated with established markers of biologically aggressive bladder cancer such as lymphovascular invasion, pathologic stage, and grade. UP III expression has limited prognostic value in patients with bladder TCC, but gene therapy viral vectors driven by the UP promoter would drive therapeutic gene expression in high-UP-expressing TCC cells, but not in aggressive low-UP-expressing TCC cells.

MP-16.02: The prognostic value of cyclooxygenase-2 expression and microvessel density in transitional cell carcinoma of the upper urinary tract

Cyclooxygenase (COX)-2 plays an important role in cancer progression by affecting tumor cell proliferation, resistance to apoptosis, angiogenesis, immune suppression, and tumor invasion in several types of cancers. However, studies about COX-2 expression in transitional cell carcinoma of the upper urinary tract (TCC-UUT) have been limited. In this study, the relationship between COX-2 expression and microvessel density (MVD), a reflection of angiogenesis with prognosis in patients with TCC-UUT, was investigated.

Clusterin Expression and Apoptosis in Transitional Cell Carcinoma of the Bladder

Purpose: The clusterin expression has been associated with tumorigenesis of various malignancies, including tumors of the prostate, colon and breast. Furthermore, the expression of clusterin is modulated by many factors that are believed to regulate tumor growth and apoptosis. We studied the clusterin expression in transitional cell carcinoma (TCC) of the bladder and we investigated its correlation with apoptosis. Materials and Methods: Eighty five bladder tumor specimens from radical cystectomy or transurethral resection were subjected to immunohistochemical clusterin staining with Ig G clusterin Ab. We examined the immunohistochemical localization of clusterin, and this was followed by TUNEL staining to detect the apoptotic cells. After double-staining with Hoechst 33258, we detected the apoptotic cells under a fluorescence microscope and we calculated the apoptotic index. Results: Invasive TCC showed a stronger positive expression of clusterin as compared with superficial TCC, but the positivity of the clusterin expression was not in proportion to the tumor grade. The apoptotic indices of cancer were 0.52±0.28%, 0.30±0.16% and 0.17±0.11% in Grade I, Grade II and Grade III superficial TCC, respectively, and it was 0.23± 0.13% in Grade III invasive TCC. Apoptotic cells were not detected in the cancer cells stained with clusterin. Conversely, clusterin was not expressed in the cells showing apoptosis. Conclusions: These results suggest that clusterin could be used as a marker to provide prognostic information for the TCC patients. The apoptotic index revealed that apoptosis and the clusterin expression have correlation with transitional cell cancer. Further study will be needed to clarify the role of clusterin as a therapeutic target for cancer treatment. (Korean J Urol 2007;48:402-407)

Increased expression of the polycomb group gene, EZH2, in transitional cell carcinoma of the bladder: Raman JD, Mongan NP, Tickoo SK, Boorjian SA, Scherr DS, Gudas LJ, Department of Urology, The New York Presbyterian Hospital, Weill Medical College of Cornell University, New York, NY

The Polycomb group gene, EZH2, functions as a transcriptional repressor involved in gene silencing. Amplification of EZH2 has been reported in several malignancies, including prostate, breast, and lymphoma. We evaluated EZH2 mRNA and protein expression in bladder specimens from patients and the EZH2 mRNA expression in five bladder cancer cell lines. EZH2 mRNA expression was assessed by reverse transcription-PCR (RT-PCR) in 38 bladder tissue specimens. We also evaluated 39 bladder cancer specimens for EZH2 protein expression using immunohistochemistry with affinity-purified antibodies to human EZH2. In addition, five human bladder cancer cell lines were analyzed by RT-PCR for EZH2 mRNA expression. Five of 14 (36%) nontumor bladder specimens versus 21 of 24 (88%) bladder tumors showed EZH2 mRNA expression (P = 0.003). All of the invasive tumors (10 of 10) had detectable EZH2 mRNA expression, compared with 11 of 14 (79%) superficial tumors. In addition, EZH2 mRNA expression was noted in 100% (16 of 16) of high-grade bladder tumors versus 50% (4 of 8) of low-grade tumors (P = 0.01). EZH2 protein expression, meanwhile, was increased in neoplastic tissue compared with nontumor urothelium (78% versus 69% of nuclei, P < 0.005). There were no differences in EZH2 protein levels between superficial and invasive tumors. High-grade tumors had increased EZH2 staining compared with normal urothelium (78% versus 68%, P < 0.005), whereas low-grade lesions did not. Four of five human bladder cancer cell lines expressed high levels of EZH2, whereas only low levels were detected in one cell line. We report a significant increase in EZH2 expression in transitional cell carcinoma of the bladder compared with normal urothelium. These data suggest that similar to other human malignancies, increased EZH2 expression correlates with oncogenesis of the bladder.

Increased Expression of the Polycomb Group Gene, EZH2, in Transitional Cell Carcinoma of the Bladder

The Polycomb group gene, EZH2, functions as a transcriptional repressor involved in gene silencing. Amplification of EZH2 has been reported in several malignancies, including prostate, breast, and lymphoma. We evaluated EZH2 mRNA and protein expression in bladder specimens from patients and the EZH2 mRNA expression in five bladder cancer cell lines. EZH2 mRNA expression was assessed by reverse transcription-PCR (RT-PCR) in 38 bladder tissue specimens. We also evaluated 39 bladder cancer specimens for EZH2 protein expression using immunohistochemistry with affinity-purified antibodies to human EZH2. In addition, five human bladder cancer cell lines were analyzed by RT-PCR for EZH2 mRNA expression. Five of 14 (36%) nontumor bladder specimens versus 21 of 24 (88%) bladder tumors showed EZH2 mRNA expression (P=0.003). All of the invasive tumors (10 of 10) had detectable EZH2 mRNA expression, compared with 11 of 14 (79%) superficial tumors. In addition, EZH2 mRNA expression was noted in 100% (16 of 16) of high-grade bladder tumors versus 50% (4 of 8) of low-grade tumors (P=0.01). EZH2 protein expression, meanwhile, was increased in neoplastic tissue compared with nontumor urothelium (78% versus 69% of nuclei, P<0.005). There were no differences in EZH2 protein levels between superficial and invasive tumors. High-grade tumors had increased EZH2 staining compared with normal urothelium (78% versus 68%, P<0.005), whereas low-grade lesions did not. Four of five human bladder cancer cell lines expressed high levels of EZH2, whereas only low levels were detected in one cell line. We report a significant increase in EZH2 expression in transitional cell carcinoma of the bladder compared with normal urothelium. These data suggest that similar to other human malignancies, increased EZH2 expression correlates with oncogenesis of the bladder.

Micro-array analysis of the effect of post-transurethral bladder tumor resection urine on transforming growth factor-β1 dependent gene expression in transitional cell carcinoma ☆

Introduction and Objectives Prior studies have shown that bladder trauma occurring during transurethral bladder tumor resection increases urinary levels of the cytokine transforming growth factor (TGF)-β1. This study used complementary deoxyribonucleic acid micro-array technology to identify additional genes in human transitional cell carcinoma (TCC), whose expression is altered as a consequence of increased urinary levels of TGF-β1. Methods The human TCC line 253J was cultured in standard media, or media spiked with either 10% post-transurethral bladder tumor resection urine (PTU), or PTU and anti-TGF-β1 neutralizing antibody. Messenger ribonucleic acid from these conditions, together with messenger ribonucleic acid from stably transfected 253J cells over-expressing TGF-β1, was hybridized with ATLAS® micro-array membranes (Clontech, Palo Alto, CA) containing 588 human genes. Hybridization signal intensity was quantified using phospho-imaging. An analytic strategy based on the variance in the signal intensity ratio of specific housekeeping genes in control and experimental comparisons was used to identify significant changes in gene expression. Reverse transcriptase polymerase chain reaction of target genes was used to confirm gene over-expression and TGF-β1 responsiveness. Results Seven genes were identified on micro-array: v-RAF-1, colony stimulating factor-1 receptor, v-FGR, insulin growth factor-1 receptor, epidermal growth factor receptor, α5 integrin, and interferon receptor-1. Reverse transcriptase polymerase chain reaction confirmed over-expression in the autocrine TGF-β1 producing cell line and increased expression in response to exogenous TGF-β1. Conclusions TGF-β1 in PTU alters the expression of multiple genes in human TCC in vitro. The impact of these changes on the biologic phenotype of the malignant cell and the efficacy of adjuvant therapies requires further evaluation.

Synergistic Induction of DOC-2/DAB2 Gene Expression in Transitional Cell Carcinoma in the Presence of GATA6 and Histone Deacetylase Inhibitor

The down-regulation of DOC-2/DAB2 gene, which encodes a unique phosphoprotein modulating signal pathways elicited by exogenous stimuli, is often associated with several cancer types; however, the underlying mechanism is still unknown. Dramatically different expression levels of DOC-2/DAB2 mRNA and protein are observed among several human transitional cell carcinoma (TCC) cell lines, suggesting that transcriptional regulation may play a role in these cells. In this study, we have shown that the histone acetylation status associated with the 5' upstream regulatory sequence of DOC-2/DAB2 gene is one of the key determinants for its gene expression. In addition, GATA6 but not other GATA family members, such as GATA2 and GATA4, can specifically induce DOC-2/DAB2 promoter activity, although GATA transcription factors share a very similar DNA-binding sequence. We also show that increased histone acetylation and the presence of GATA6 have a synergistic effect on DOC-2/DAB2 promoter activity, which results in the elevation of DOC-2/DAB2 protein expression. Thus, we conclude that transcriptional regulation of DOC-2/DAB2 gene in human TCC is determined by histone acetylation and a specific transcription factor (i.e., GATA6), which underlie the reduced DOC-2/DAB2 protein expression in TCC cells.

773: Effect of Natural Compound on the Expression of Coxsackie and Adenovirus Receptor Gene Expression in Transitional Cell Carcinoma

773: Effect of Natural Compound on the Expression of Coxsackie and Adenovirus Receptor Gene Expression in Transitional Cell Carcinoma

Expression of matrix metalloproteinase-10 in human bladder transitional cell carcinoma

Objectives To analyze matrix metalloproteinase-10 (MMP-10) expression in transitional cell carcinoma (TCC) of the bladder, evaluate the correlations between MMP-10 protein expression and clinicopathologic parameters, and address the viability of MMP-10 as a therapeutic target for TCC. MMP-mediated degradation of the extracellular matrix is an important factor in the pathogenesis of tumorigenesis and metastasis. Methods Using immunohistochemistry, the expression of MMP-10 was assessed using both tissue microarrays and whole sections of archival tissue specimens representative of all grades and stages of human bladder TCC (n = 60). MMP-10 expression was also assessed in histologically normal human bladder tissue (n = 10). The immunostaining results for MMP-10 expression were examined for correlations with tumor grade and stage. Results Unlike most MMPs, MMP-10 was localized primarily in the tumor mass as opposed to the tumor stroma and was detectable in all grades and stages of TCC. Significantly greater levels of MMP-10 protein were observed in superficial (pTa, pT1; n = 38) tumors than in normal bladder tissue ( P = 0.01). In contrast to the proposed role of MMPs in tumor invasion, no significant difference was observed between muscle-invasive tumors (pT2 or worse; n = 22) and histologically normal bladder tissue ( P = 0.50). MMP-10 expression showed no significant correlation with tumor grade. Conclusions The data from our study showed that, unlike most MMPs, MMP-10 was not associated with tumor aggression or invasion. Our results suggest that MMP-10 protein levels are significantly greater in the earlier stages of TCC development.

Expression of Hypoxia-Inducible Factor-1α Protein Predicts Survival in Patients with Transitional Cell Carcinoma of the Upper Urinary Tract

Hypoxia-inducible factor-1 (HIF-1), identified as one of the transcription factors, has been found to play an essential role in oxygen homeostasis. HIF-1 is a heterodimer composed of HIF-1alpha and HIF-1beta. Increased levels of HIF-1alpha have been reported during the carcinogenesis and progress of several tumors. We investigated the prognostic importance of HIF-1alpha expression in transitional cell carcinoma of the upper urinary tract. In 127 cases of transitional cell carcinoma of the upper urinary tract, we examined its expression (using immunohistochemistry and in situ hybridization), and also its relation to the expression of p53 oncoprotein, as well as to proliferating cell nuclear antigen (PCNA) immunoreactivity, microvessel density, clinicopathologic parameters, and clinical outcome. A positive expression of HIF-1alpha protein was recognized in 55.1% of samples, the expression being apparent within the nucleus in tumor cells. HIF-1alpha protein expression correlated with grade, growth pattern, p53 oncoprotein expression, and PCNA index, but not with stage. Furthermore, a significant correlation was found between HIF-1alpha protein expression and both overall and disease-free survival rates in the univariate and multivariate analyses (in all tumors and in invasive tumors). A positive expression of HIF-1alpha mRNA was recognized in 69.6% of 125 samples which were available, the expression being apparent within the cytoplasm in tumor cells. The positive expression of HIF-1alpha mRNA by in situ hybridization correlated significantly with HIF-1alpha protein expression by immunohistochemistry. HIF-1alpha mRNA expression only correlated with pattern of growth (P = 0.0078). In conclusion, the detection of HIF-1alpha protein would seem to be of value in informing the prognosis of transitional cell carcinoma of the upper urinary tract.

APOPTOSIS AND CHEMOTHERAPY FOR BLADDER CANCER

We discuss the role of apoptosis, that is gene directed self-destruction of a cell, in the response of bladder transitional cell carcinoma cells to chemotherapy. A directed MEDLINE literature search of apoptosis, bladder cancer and chemotherapy was performed to extract relevant information for review. The characteristics of apoptotic cells were defined and the methods in common use to detect these traits is described. The role of the key mediators of the apoptotic process in bladder cancer is discussed in the context of chemosensitivity and disease stage. The importance of the apoptosis induction after chemotherapy is highlighted. On stimulus by appropriate external or internal signals a cell may alter the expression of genes encoding for proteins associated with the apoptotic process. The development of apoptosis depends on the balance between pro-apoptotic and anti-apoptotic proteins. Key alterations in genes and proteins related to apoptosis within bladder cancer result in a shift away from the default state of apoptosis toward a cell with increased survival properties that is chemoresistant. Much current research in bladder cancer is aimed at restoring chemosensitivity by shifting the cell toward a pro-apoptotic phenotype. Successful translation of this work into clinical practice may improve survival in patients in whom prognosis is currently poor.