The expression of oncoproteins in transitional cell carcinoma: its correlation with pathological behavior,cell cycle and drug resistance

OBJECTIVE: To investigate the relationship of oncoproteins with histological grade, tumor stage, cell cycle and multidrug resistance (MDR) in bladder cancer. METHODS: The expression of various oncoproteins (p21, EGFR1, erbB-2, c-jun, c-myc, bcl-2, Bax) and suppressor gene products (WT-1, RB gene, p53) was studied in normal urothelium, transitional cell carcinoma (TCC) cell lines and their MDR sublines by using specific antibodies and an indirect immunofluorescence flow-cytometric method. RESULTS: The total increased rate of measured oncoproteins in TCC cell lines was 62.7%. There was no difference in the expression rate of oncoproteins between low-grade/stage TCC cell lines and high-grade/stage TCC cell lines. All of these TCC cell lines have aneuploidy and increased proliferative activity in the cell cycle, but a correlation with oncoprotein expression was not seen. They had a low expression rate of c-myc, Bax and RB gene when compared to normal urothelium. On the contrary, the expressions of p21, EGFR1, erbB-2, bcl-2, WT-1 and p53 oncoproteins were significantly higher than in normal urothelium (p < 0.05). A long-term cultured MDR subline of TCC8702 (TCC8702/ADR1000) demonstrated a generally decreasing expression of oncoproteins in addition to p53. CONCLUSIONS: The p21, EGFR1, erbB-2, bcl-2, WT-1 and p53 oncoproteins were increased in TCC cells compared to normal urothelium. Oncoprotein expression is related to tumorigenesis of TCC cells, but no correlation was seen between the incidence and tumor differentiation and cell cycle. These oncoproteins decreased gradually in the process of generation of MDR in addition to p53. It indicates that the p53 oncogene is closely related to the acquired MDR of TCC cells.     

The expression of vascular endothelial growth factor in transitional cell carcinoma of urinary bladder is correlated with cancer progression

Vascular epithelial growth factor (VEGF) regulates neovascualrization in malignant cells. VEGF as a mitogen is thought to alter cancer cell formation and tumor progression. We aimed to investigative the expression of the VEGF gene to evaluate their clinical significance in transitional cell carcinoma (TCC) of urinary bladder. Tissue samples from 161 patients with TCC were examined with an immunohistochemical stain for the expression of the VEGF gene. The expression rate was compared to 32 normal bladder mucosal samples obtained from transurehtral surgery from noncancer patients. The results revealed significant differences between normal urothelium (0%) and cancer tissue (54.7%) for the positive staining of VEGF protein (P < 0.001). With the progression of tumor grade and clinical staging, the positive rate of VEGF gene expression significantly increased. Expression of the VEGF gene in the invasive group was greater than that in the noninvasive group (P < 0.001). The results revealed that expression of the VEGF gene is proportional to the formation and progression of TCC. Therefore, abnormal expression of VEGF genes can be used as a prognostic marker in TCC of urinary bladder.     

Transient receptor potential vanilloid type 1 (TRPV1) expression changes from normal urothelium to transitional cell carcinoma of human bladder

OBJECTIVE: To investigate possible changes in the expression of the transient receptor potential vanilloid type 1 (TRPV1) from normal urothelium to transitional cell carcinoma (TCC) of human bladder. METHODS: Specimens from normal bladder (n=13, mean age 62 yrs), superficial TCC (n=16, mean age 62,4 yrs) and muscle invasive bladder cancer (n=12, mean age 67 yrs), were obtained by multiple cold cup and full-thickness biopsy during open surgery. All the specimens were processed for H&E staining, immunohistochemistry and Western Blot analysis. RESULTS: In controls, the urothelium showed a labelling whose intensity was higher in the superficial cells than in the basal and club-shaped ones. In the superficial TCC, the urothelium showed a reduced labelling intensity. In the muscle invasive TCC, a very light labelling was occasionally detected in scattered superficial cells and no labelling was present in the basal cells and in those that had invaded the muscle. In controls, Western Blot analysis recognized two thick, intensely stained bands, with a molecular weight of approximately 100 and 95 kDa. In all superficial TCC there were two bands similar to control ones and in the muscle invasive two very thin, lightly stained bands. No band was detected in the patients staged as pT4. CONCLUSION: These data demonstrated a progressive loss of TRPV1 expression in the urothelium as TCC stage increased and cell differentiation was lower. Future studies will establish the importance of this loss for TCC differentiation and progression.     

Tumor recurrence in the remnant urothelium of females undergoing radical cystectomy for transitional cell carcinoma of the bladder: long-term results from a single center

PURPOSE: We analyzed the risk factors and incidence of secondary TCC of the remnant urothelium in women following radical cystectomy for TCC of the bladder. MATERIALS AND METHODS: A total of 85 women with a mean age of 64.5 years with clinically localized TCC of the bladder underwent radical cystectomy between 1992 and 2004. Orthotopic bladder substitution was performed in 46 females, while 39 underwent nonorthotopic urinary diversion. Of the entire cohort 22 (26%) patients underwent cystectomy for multifocal or recurrent TCC. Followup examinations were performed at 6-month intervals. RESULTS: Mean followup in the entire cohort was 49.8 months (median 42). Intraoperative frozen sections obtained from the urethra and distal ureters were negative for TCC and CIS in all patients. Four women (4.7%) had TCC in the remnant urothelium at a mean of 56 months postoperatively. These patients had undergone cystectomy for multifocal or recurrent TCC (4 of 22 or 18%). No secondary TCC was seen in the 63 patients with solitary invasive or nonrecurrent bladder cancer (p <0.05). Urethral recurrence was found in 2 patients (4.3%) 65 and 36 months after orthotopic neobladder surgery, respectively. In the orthotopic group 1 patient (2.1%) had an upper urinary tract tumor 76 months after surgery, while in the nonorthotopic group 1 (2.5%) was found to have an upper urinary tract tumor 48 months postoperatively. CONCLUSIONS: Recurrent or multifocal TCC may represent a risk factor for secondary TCC of the remnant urothelium after cystectomy. In our series all recurrent tumors were late recurrences (more than 36 months postoperatively). Because the rate of urethral recurrence in the current series corresponds to that reported in men (2% to 6%), urethra sparing cystectomy with orthotopic bladder replacement does not appear to compromise the oncological outcome in women.     

Clinical relevance of maspin expression in bladder cancer

Transitional cell carcinoma (TCC) of the bladder is a solid tumor that induces angiogenesis to maintain nutrition and oxygenation of tumor cells. Maspin, a serpin with tumor suppressing activity, has recently been identified as an inhibitor of angiogenesis. This study examined the impact of maspin expression in the growth pattern of TCC of the bladder. Maspin was identified in a panel of normal tissues, in several bladder carcinoma cell lines, and 51 patient samples of TCC of the bladder. Expression was detected by RT-PCR and immunohistochemistry. Furthermore, the level of maspin was correlated to the growth rate of bladder tumor cell lines in vitro and in vivo. Maspin expression was found in high quantities in normal urothelium. Maspin expression was preserved in superficial bladder cancers but was significantly diminished in invasive carcinomas. Within the group of invasive TCCs, maspin expression was inversely correlated to the patient prognosis. Furthermore, low maspin expression level was coupled to an increased tumor cell growth in vivo. Down-regulation of maspin expression seems to be a specific event in the progression of invasive bladder carcinoma. Maspin might be a useful marker to determine the prognosis of invasive TCC. Furthermore, maspin re-expression might become a therapeutic option in the treatment of invasive, metastatic TCC.This study was supported in part by Medac GmbH.     

Histopathological and immunohistochemical study of papillary urothelial neoplasms of low malignant potential and grade associated with extensive invasive low-grade urothelial carcinoma

OBJECTIVE: To report five cases of papillary urothelial neoplasm of low malignant potential (UNLMP) and papillary urothelial carcinoma of low grade (UCLG) associated with extensive muscle invasion, and to investigate the clinical and histopathological presentation and their immunohistochemical properties. MATERIALS AND METHODS: Consecutive cystectomy and correlating transurethral resection (TUR) of urinary bladder tumour specimens were reviewed to identify cases of UCLG having extensive invasion into the urinary bladder wall. All specimens were stained immunohistochemically, as were those from 10 control cases having reactive urothelium or superficial UNLMP. The clinical charts were reviewed. RESULTS: Of a total of 95 cystectomy cases there were four of UNLMP or UCLG with extensive invasion. An additional case was added from our consultation file. All five cases had biopsies misdiagnosed as benign lesions or prostatic adenocarcinoma. The superficial invasive components consisted of UCLG conforming to the previously described entities of nested transitional cell carcinoma (TCC), microcystic or deceptively benign-appearing TCC. Immunostaining for cytokeratin 20, MIB-1 and p53 was similar to reactive epithelia, whereas E-cadherin immunoreactivity was slightly different, with focal negativity compared with extensive immunoreactivity in invasive vs noninvasive UCLG. Four patients developed distant metastases; three died within a follow-up of 3 years. CONCLUSIONS: UNLMP and UCLG that widely and deeply invade the bladder accounted for 4% of urothelial carcinoma (UC) in cystectomy specimens and commonly pose diagnostic problems in superficial TUR specimens. From this study with few cases the diagnosis of this entity in superficial biopsies is aided by an awareness of it and by identifying 'benign appearing' nests of urothelial cells which are deeply seated in the stroma. Immunostaining is unlikely to be very useful.     

膀胱移行细胞癌组织中黑色素瘤抗原基因及蛋白的表达

目的探讨膀胱移行细胞癌(TCC)中黑色素瘤抗原(MAGE)基因mRNA的表达及其临床意义。方法采用逆转录聚合酶链反应(RT PCR)技术检测3个膀胱TCC细胞株和20例膀胱TCC患者癌组织(新鲜标本,T1期7例,T2期5例,T3期6例,T4期2例;G11例,G211例,G38例)MAGE A1,A2,A3,A4mRNA表达,免疫组化法检测105例膀胱TCC患者癌组织(石蜡标本,T1期35例,T2期12例,T3期26例,T4期13例,另19例无法确定分期;G113例,G244例,G348例)MAGE A4蛋白的表达。结果3个膀胱TCC细胞株均有MAGE基因mRNA的表达;20例膀胱TCC新鲜标本组织MAGEmRNA阳性:A112例(60%),A216例(80%),A311例(55%),A418例(90%),A1～A4均阳性8例(40%)。105例膀胱TCC石蜡标本组织中MAGE A4蛋白阳性53例(50%),高分级膀胱TCC组织中强表达(++或+++)率为27%(13/48),显著高于低分级的4%(2/57)(F=12.00,P<0.01),高分期膀胱TCC组织中强表达率为27%(14/51),显著高于低分期的0%(0/35)(F=11.48,P<0.01)。结论MAGE基因在膀胱TCC中有较高表达,分级或分期高的膀胱TCC中MACE A4蛋白明显强表达。     

Differential Expression of Hepatocyte Growth Factor in Papillary and Nodular Tumors of the Bladder

Background: The aim of this study was to investigate the expression of hepatocyte growth factor (HGF) and its receptor (c-met) during bladder tumorigenesis.
Methods: HGF and c-met expression were analyzed immunohistochemically in matched samples of normal, dysplastic, and carcinoma specimens from 49 human bladders resected at the time of radical cystectomy for nonmetastatic transitional cell carcinoma (TCC). The tumors were composed of papillary (n = 22), nodular (n = 16) or mixed papillary and nodular (mixed; n = 11) components. Results: The normal urothelium showed no significant immunoreactivity to HGF. Expression of HGF was observed in 45.5%, 77.3% and 90.9% of specimens demonstrating mild, moderate, and severe dysplastic lesions adjacent to papillary TCCs, respectively, whereas all of the papillary TCC samples were positive for HGF. No immunoreactivity for HGF was found in dysplastic lesions from nodular tumors, and only 2 specimens had positive immunostainingfor HGF in the tumor areas (1 showed weak immunostainingand 1 showed HGF immunostainingonly in the deeper invasive compartment). Additionally, 3 nodular lesions taken from mixed tumors showed weak immunostaining for HGF while the concurrent papillary lesions were HGF-positive. There was a significant difference of HGF immunoreactivity between papillary and nodular tumors (P < 0.01). c-met immunostaining was consistently detected in all specimens. HGF and c-met immunoreactivity did not significantly correlate with tumor stage and grade, nor with overall patient survival irrespective of the tumor growth pattern.
Conclusion: These results suggest that HGF expression may play a significant role in the development of papillary TCC.     

Upper urinary tract tumour after radical cystectomy for transitional cell carcinoma of the bladder: an update on the risk factors, surveillance regimens and treatments

Urothelial carcinoma is characterized by multiple, multifocal recurrences throughout the genitourinary tract; approximately 3% of patients treated by radical cystectomy (RC) for invasive transitional cell carcinoma (TCC) of the bladder will subsequently develop a subsequent TCC in the upper urinary tract (UUT) urothelium. Metachronous upper UUT tumours (mUUT-TCC) typically occur as a late oncological event (>3 years after RC). The vast majority of mUUT-TCCs are detected only after the progression to tumour-related symptoms, e.g. haematuria, flank pain or pyelonephritis, despite strict adherence to surveillance protocols. Failure of imaging and cytology to detect most asymptomatic tumours has led to questions about the need for routine UUT surveillance. Some authors have advocated a more tailored approach to surveillance after RC, targeting high-risk patients and with limiting imaging in those patients at lowest risk of developing a subsequent UUT-TCC. mUUT-TCCs are most common in patients with TCC in the ureter or urethra, and with organ-confined bladder cancer. Although the prognosis is generally poor, long-term survival can be achieved in a subset of patients after radical nephroureterectomy (NU). Minimally invasive techniques, e.g. ureteroscopic and percutaneous resection, have been proposed as renal-sparing alternatives to radical surgery for patients with low-stage and -grade de novo UUT-TCC. However, oncological control of renal-sparing therapies in those with high-risk mUUT-TCC remains largely unconfirmed. Until oncological outcomes equivalent to the standard, radical NU, are reported in patients after RC, conservative treatment strategies should be avoided.     

DNA hypermethylation on multiple CpG islands associated with increased DNA methyltransferase DNMT1 protein expression during multistage urothelial carcinogenesis

PURPOSE: We elucidated the significance of aberrant DNA methylation on multiple CpG islands and its correlation with DNA methyltransferase DNMT1 protein expression during urothelial carcinogenesis. MATERIALS AND METHODS: We examined the DNA methylation status on multiple CpG islands by methylation specific polymerase chain reaction and combined bisulfite restriction enzyme analysis in 12 specimens of normal urothelium, 23 of noncancerous urothelium showing no remarkable histological changes obtained from patients with bladder cancer (NBC) and 70 of transitional cell carcinoma (TCC). RESULTS: DNA methylation on CpG islands of the p16 (0%, 17% and 21%) and death-associated protein kinase (13%, 33% and 29%) genes, and methylated in tumor-2 (56%, 60% and 76%), 12 (0%, 6% and 30%), 25 (25%, 27% and 35%) and 31 (45%, 56% and 79%) clones was detected in normal urothelium, NBCs and TCCs, respectively. The incidence of concurrent DNA hypermethylation on 3 or more CpG islands in NBCs (38%) was significantly higher than that in normal urothelium (0%, p = 0.0455) and even higher in TCCs (59%, p = 0.0043). The incidence of the CpG island methylator phenotype in nonpapillary carcinomas (nodular invasive carcinomas and their precursors, ie flat carcinoma in situ, 71%) was significantly higher than in papillary carcinomas (40%, p = 0.0143). In all specimens examined concurrent DNA hypermethylation on 3 or more CpG islands significantly correlated with immunohistochemically evaluated DNMT1 protein over expression (p = 0.0167). CONCLUSIONS: DNA hypermethylation on multiple CpG islands in association with DNMT1 protein over expression may participate in multistage urothelial carcinogenesis even at the precancerous stage and particularly in the development of nodular invasive carcinomas of the bladder.     

Mucin gene expression in human urothelium and in intestinal segments transposed into the urinary tract

PURPOSE: The repertoire of mucin (MUC) gene expression in the normal human urothelium is poorly defined and the alterations in MUC gene expression following transposition of intestinal segments into the urinary tract has not previously been studied. The aims of this study were to define MUC gene expression in the normal human urothelium; and in transposed intestinal segments. MATERIALS AND METHODS: Non-isotopic in-situ hybridization was carried out using eight digoxigenin labeled oligonucleotide mucin gene probes (MUC 1 - 7). Immunohistochemistry using NCL-MUC1 and NCL-MUC2 monoclonal antibodies was performed on sections of paraffin-embedded tissues. Twenty-seven patients were investigated (normal human urothelium, n = 6; transposed ileal segments, n = 14 and normal ileal controls, n = 7). RESULTS: MUC1 and MUC4 were the predominant mucin genes expressed in the normal urothelium with MUC3 being expressed in a third of cases studied; MUC2, 5AC, 5B, 6 and 7 were not expressed. Despite the morphological changes seen in transposed ileal segments, MUC2 and MUC3 continued to be expressed in these segments albeit in a disorganised fashion. Both MUC1 and MUC4 were up-regulated in transposed ileal segments, genes expressed by the normal human urothelium. All eight mucin genes were expressed in an area of pyloric-type metaplasia found in one transposed ileal segment. In patients with clam enterocystoplasty there was evidence of increasing up-regulation of MUC2, 3, 4 and 5AC expression in the urothelium toward the anastomotic site. CONCLUSION: Transposition of ileal segments into the urinary tract results in up-regulation of MUC1 and MUC4, the predominant MUC genes expressed in the human bladder. The clinical implication of the up-regulation of some MUC genes toward the anastomotic site in patients with an enteroplasty and the aberrant expression of MUC5AC - MUC7 by transposed segments is at present unclear.     

Study on nucleolar organizer regions in bladder neoplasm

Quantification of the argyrophilic nucleolar organizer regions (AgNORs), using a one-step silver colloid method, was made in proliferative urothelial lesions including neoplasm of the rat urinary bladder induced by 0.05% N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) and of the human urinary bladder. The mean numbers of AgNORs in various experimental lesions in rats given BBN for 5-30 weeks in drinking water were: nontreated urothelium (n = 6), 1.26 +/- 0.09; urothelium outside focal lesions (n = 10), 1.75 +/- 0.10; simple hyperplasia (n = 10), 2.01 +/- 0.15; papillary or nodular (PN) hyperplasia (n = 10), 2.15 +/- 0.19; papilloma (n = 5), 2.37 +/- 0.12; and transitional cell carcinoma (TCC) (n = 5), 3.52 +/- 0.23. There was a significant difference in the mean number of AgNORs between any two groups (p less than 0.05), except for simple hyperplasia vs PN hyperplasia (p = 0.085). In various proliferative lesions of human biopsy and resected samples, the mean numbers of AgNORs were normal urothelium (n = 8), 1.71 +/- 0.08; proliferative cystitis (n = 14), 1.79 +/- 0.18; hyperplasia (n = 8), 1.76 +/- 0.23; mild dysplasia (n = 5), 2.15 +/- 0.36; moderate dysplasia (n = 13), 2.61 +/- 0.27; severe dysplasia (n = 6), 3.46 +/- 0.57; G0 of TCC (n = 9), 1.95 +/- 0.17; G1 of TCC (n = 16), 2.39 +/- 0.20; G2 of TCC (n = 21), 3.33 +/- 0.31; G3 of TCC (n = 15), 4.68 +/- 0.51; and carcinoma in situ of TCC (n = 10), 3.61 +/- 0.52.(ABSTRACT TRUNCATED AT 250 WORDS)     

EXPRESSION OF BCL-2 AND BCL-X IN BLADDER CANCER

Purpose

TP53 and RB1 gene mutations in bladder transitional cell carcinoma (TCC) are correlated with grade, stage, recurrence, and survival and may correlate with tumor cell apoptotic potential. Overexpression of the bcl-2 and bcl-X anti-apoptotic genes has been correlated with poor prognosis and chemotherapy resistance in other systems. Similar studies have not been performed in TCC. We thus sought to determine expression of bcl-2 and bcl-X in TCC and correlate these with stage, survival and abnormal pRb or p53 expression.

Materials and Methods

Forty-two TCC samples (19 Ta and 23 locally advanced tumors) and normal urothelial controls were examined. Immunohistochemistry for p53, pRb, bcl-2 and bcl-X was performed on an automated system using indirect streptavidin biotin/horseradish peroxidase staining. Western immunoblot analysis was performed on bladder cancer cell lines to further characterize bcl-X expression. Recurrence-free and disease-specific survival were retrospectively determined. Kaplan-Meier survival curves were compared using the log rank test, and correlation of abnormal staining with stage and p53 or pRb status was determined using Fisher's exact test.

Results

Bcl-2 was expressed in less than 1% of normal urothelial cells, but moderate expression of bcl-x was found in all normal urothelial samples. Only 7.0% of TCC samples (1/19 Ta and 2/23 locally advanced tumors) demonstrated bcl-2 overexpression. Bcl-X overexpression was observed in 45.2% of TCC (8/19 Ta and 11/23 locally advanced tumors). Western blot analysis also revealed that both the long (29 kDa) anti-apoptotic form and short (19 kDa) pro-apoptotic form were overexpressed in bladder cancer cell lines and normal human urothelial cells. Bcl-X overexpression was weakly correlated with normal p53 expression (p = 0.06). There were no correlations of bcl-2 and bcl-X overexpression with abnormal p53, pRb, or tumor stage. There were no differences in recurrence-free or overall survival in patients with abnormal bcl-X staining.

Conclusions

Bcl-2 overexpression is rare in TCC. Bcl-X overexpression is common, likely reflecting its expression pattern in normal urothelium, but is not correlated with stage or abnormal p53 or pRb staining. Within the power limitations of this small study, bcl-X overexpression is not correlated with recurrence or survival.     

膀胱移行细胞癌表皮生长因子受体基因扩增及其表达

运用免疫组织化学技术和分子杂交技术研究了膀胱移行细胞癌表皮生长因子受体（ＥＧＦｒ）表达状况以及ＥＧＦｒ基因扩增和其ｍＲＮＡ与受体蛋白表达的相关性。结果表明：４例正常膀胱粘膜ＥＧＦｒ染色阴性；５６例膀胱肿瘤中，２９例呈不同程度的阳性染色，并与肿瘤的分期、分级有显著性差异；１３例膀胱肿瘤中，ＥＧＦｒ基因未见扩增，５例ｍＲＮＡ过表达，７例ＥＧＦｒ过表达，但二者表达的量并无相关性。提示ＥＧＦｒ表达可作为膀     

A histopathologic investigation of PGE(2) pathways as predictors of proliferation and invasion in urothelial carcinomas of the bladder

INTRODUCTION AND OBJECTIVES: The pattern of arachidonate acid (AA) transformation in tumor cells has been shown to play a role in determining tumor cell invasiveness. AA is released from membrane phospholipids by cPLA(2). Then it is metabolized into prostaglandins and PGE(2) especially via cyclooxygenase pathways. PGE(2) production seems to be necessary for rendering the cells invasive. We aimed to characterize cPLA(2), cyclooxygenase 2 (COX2) and prostaglandine E synthase (PGES) expression in human transitional carcinoma (TCC) of the urinary bladder and correlate with the Ki-67 proliferating marker. METHODS: Formalin-fixed human TCC tissues (n=54) obtained from TURB or cystectomies were evaluated for cPLA(2), COX2, PGES and Ki-67 expression using specific antibodies. There were 6 CIS, 9 pTaG1, 9 pTaG3, 10 pT1G3 and 10 pT2G3. 10 normal bladder tissues were also evaluated. Control slides were incubated without primary antibodies and treated in a similar way. RESULTS: cPLA(2), COX2 and PGES were not expressed in the 10 normal tissues. In the same normal tissues, Ki-67 expression was observed only in 1% of the cells. However, cPLA(2) was expressed in 1/6 CIS, 1/9 pTaG1, 3/9 pTaG3, 6/10 pT1G3 and 2/10 pT2G3. COX2 was expressed in 0/6 CIS, 0/10 pTaG1, 2/9 pTaG3, 3/10 pT1G3 and 1/10 pT2G3. PGES was expressed in 4/6 CIS, 0/9 pTaG1, 4/9 pTaG3, 2/10 pT1G3 and 5/10 pT2G3. Ki-67 expression was 39.5% for CIS, 6.5% in pTaG1, 37% in pTaG3, 34.5% in pT1G3 and 55% in pT2G3. If we consider it a positive result when at least one enzyme was expressed, there were 5/6 CIS positive, 1/9 pTaG1 positive, 9/9 pTaG3 positive, 10/10 pT1G3 positive and 10/10 pT2G3 positive. Also the Ki-67 is more often expressed in cells with high grade tumor. CONCLUSIONS: These results suggest that (i). not only COX2 is involved in the tumorogenesis of the TCC but also cPLA(2) and PGES, (ii). there is relationship between the AA metabolic PGE(2) pathway expression and the aggressiveness of the TCC of the urinary bladder.     

Chorionic gonadotropin beta-subunit and core fragment in bladder cancer: mRNA and protein expression in urine,serum and tissue

OBJECTIVES: Many transitional cell carcinomas (TCC) of the bladder express the beta-subunit (CGbeta) of chorionic gonadotropin (CG), and elevated serum levels occur especially in advanced disease. We have compared the diagnostic utility of various methods for detecting CG and CGbeta expression at the protein and mRNA level. METHODS: We used RT-PCR to detect CGbeta mRNA in urinary cells and highly sensitive immunoassays to determine CG and CGbeta in serum and the core fragment of CGbeta (CGbetacf) in urine from patients under follow-up for bladder cancer. Tissue expression was studied by immunohistochemistry. RESULTS: CGbeta mRNA was detected in urinary cells in 50% (n=84) of the cancer cases and in none of the healthy controls (n=15). Positive staining for CGbeta in tissue samples was observed not only in 30% (n=96) of the TCC cases, but also in 5 of 20 histologically benign samples from TCC patients, and in 10 of 21 samples from benign bladder diseases. Serum and urinary concentrations of CGbeta were elevated in 29% (n=66) and 8% (n=72), respectively, while serum CG was elevated in 18% of the TCC patients. Urinary CGbetacf concentrations were higher in invasive (T1-T4) than superficial (T in situ and Ta) tumors (p=0.037), in cases positive for CGbeta mRNA (p=0.03) and cases with suspicious or malignant urinary cytology (p=0.002). The ratio of urinary to serum concentration of CGbeta showed the strongest correlation with tumor stage (p<0.00001), grade (p<0.00001), and staining for CGbeta (p=0.019). CONCLUSIONS: Although CGbeta expression may occur in benign bladder epithelium, CGbeta mRNA in urinary cells is a potential marker of bladder cancer. Urinary and serum CGbeta have low sensitivity in early disease, but the urine/serum ratio appears to indicate local release of CGbeta into urine. Further studies are needed to evaluate the clinical usefulness of different forms of CGbeta expression.     

Urinary levels of soluble e-cadherin in the detection of transitional cell carcinoma of the urinary bladder

OBJECTIVE: To test the hypothesis that elevated urinary levels of soluble E-cadherin (sE-cadherin) would aid in the detection of transitional cell carcinoma (TCC) of the urinary bladder. METHODS: We performed sE-cadherin staining of one murine (MBT2) and four human (RT4, 5637, T24, and TCCSUP) bladder cancer cell lines. sE-cadherin levels were also determined in voided urine of 188 consecutive subjects at risk for TCC recurrence, 31 patients with other uro-pathologic conditions, and 10 healthy subjects using a commercially-available ELISA kit. sE-cadherin was analyzed continuously and categorically on the basis of its median distribution. RESULTS: Moderately and poorly differentiated bladder cancer cell lines had decreased cellular E-cadherin expression, whereas RT4, a well differentiated cell line, had preserved expression. All cell lines had measurable sE-cadherin levels in their conditioned media. The area under the ROC curve of sE-cadherin for the detection of TCC was 0.719 (95%CI, 0.637-0.801; p<0.001). Higher levels of sE-cadherin were associated with positive cytology results (p=0.012) and muscle invasive tumor stage (p=0.009). Urinary sE-cadherin was more sensitive, but less specific than urinary cytology for the detection of bladder TCC. In a multivariable logistic regression analysis, higher sE-cadherin and positive cytology were both associated with an increased risk of bladder TCC (p=0.048 and p<0.001, respectively). Combination of cytology and sE-cadherin allowed categorization of patients into three significantly different risk groups for bladder cancer. Adjustment of sE-cadherin for urinary creatinine levels did not affect any of the outcomes. CONCLUSIONS: Urinary level of sE-cadherin may add information to cytology in the detection of bladder TCC.