Expression of p53 product in Chinese human bladder carcinoma

Summary Expression of p53 protein was examined in 67 cases of primary transitional cell carcinoma of the bladder and 6 normal controls using an immunohistochemical method on paraffin sections. Positive nuclear staining for p53 in malignant cells was found in 34 (51%) of the 67 cancer patients; no positive staining for p53 was detected in any of the normal controls or in the benign cells, including stromal and inflammatory cells, within the tumor tissue. There were 8 positive cases (33%) in 24 grade G1 tumors, 12 (48%) in 25 G2 tumors and 14 (78%) in 18 G3 tumors. p53 protein was detected positively in 14 (36%) of 39 superficial tumors (Tis-T1) and in 20 (71%) of 28 invasive tumors (T2–T4). Thus, positive staining for p53 was found more frequently in poorly differentiated tumors (chi-squared test: G3/G1+G2P<0.01) and in invasive tumors (chi-squared test: T2–T4/Tis-T1P<0.01). Expression of p53 was also closely associated with recurrence of tumors. Alterations in p53 expression may be of prognostic value in cases of bladder transitional cell carcinoma.     

The significance of p53 mutations as an indicator of the biological behavior of recurrent hepatocellular carcinomas

The significance of p53 mutations in the primary lesion for recurrent hepatocellular carcinoma (HCC) was evaluated. Mutations of p53 were examined using non-radioisotopic (nonRI)-polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP) in 98 resected HCCs. Of the 98 cases, 25 (26%) had a p53 mutation. In 83 patients who survived surgery, the presence of a p53 mutation was associated with a shortened overall survival (P<0.001) and a shortened cancer-free survival (P<0.05). In 43 patients who developed recurrence, there was no statistically significant correlation between the status of p53 in the primary lesion and the clinical features of recurrent HCCs examined, i.e., extrahepatic metastasis, the number of recurrent tumors, extent of recurrent tumors, and treatment for recurrent tumors. However, postrecurrence survival was significantly lower in patients in whom a p53 mutation had been detected in the primary lesion (P<0.01). A multivariate analysis for prognostic value after recurrence revealed that the p53 mutation was a useful independent prognostic factor affecting survival after recurrence (P<0.01). In conclusion, our findings suggest that HCCs with p53 mutations have a high malignant potential based on their poor prognosis. Therefore, a p53 mutation in the primary lesion is useful as an indicator of the biological behavior of recurrent HCCs.     

p53 expression in patients with advanced urothelial cancer of the urinary bladder

Study Type – Prognosis (inception cohort)
Level of Evidence 1b

OBJECTIVE

To test whether assessing p53 expression could improve the ability to predict disease recurrence and disease‐specific survival in a multi‐institutional cohort of patients with advanced urothelial carcinoma of the urinary bladder (UCB).

PATIENTS AND METHODS

The study comprised 692 patients with pT3–4 N0 or pTany N+ UCB treated with radical cystectomy and lymphadenectomy. The predictive accuracy (PA) was quantified using the 200 bootstrap‐corrected concordance index. The base model comprised age, gender, stage, grade, lymphovascular invasion, number of lymph nodes removed, number of lymph nodes positive, concomitant carcinoma in situ, and adjuvant chemotherapy.

RESULTS

p53 expression was altered in 341 (49.3%) patients. In multivariable analyses, p53 expression was independently associated with disease recurrence (hazard ratio, 1.66; P < 0.001) and cancer‐specific mortality (hazard ratio 1.65, P < 0.001). Overall, adding p53 did not significantly improve the PA of the base model (recurrence +0.7%, P = 0.085, and cancer‐specific mortality +1.2%, P = 0.050). In the subgroups of pT3N0 (280) and pT4N0 (83) patients, p53 slightly improved the PA of the base model by a statistically significant degree (recurrence +1.7% and +3.6%, respectively; cancer‐specific mortality +1.9% and +3.5%, respectively; all P < 0.001). In 329 patients with pTany N+ disease p53 status did not improve the PA of the base model.

CONCLUSION

While assessing p53 expression has limited utility in patients with lymph node‐positive UCB, it marginally improves prognostication in patients with advanced non‐metastatic UCB. Integration of p53 into a panel of biomarkers might be necessary to capture a more accurate picture of the biological potential of advanced UCB.     

p130Cas,E‐cadherin and β‐catenin in human transitional cell carcinoma of the bladder: Expression and clinicopathological significance

Objectives: To investigate the effects of junction protein, p130 Crk‐associated substance (p130Cas), and adhesion molecules, E‐cadherin and β‐catenin, on the biological behavior of transitional cell carcinoma of the bladder. Methods: In 72 paraffin embedded specimens of transitional cell carcinoma of the bladder and 20 normal controls, the expression of p130Cas, E‐cadherin and β‐catenin was examined by quantum dot‐based immunofluorescence histochemistry (QD‐IHC) and conventional immunohistochemistry (IHC). Results: QD‐IHC was consistent with IHC in detecting the expression of the three molecules (P > 0.05 for all comparisons). The positive expression rate of p130Cas in bladder cancer tissues increased more significantly than that in normal bladder tissues (P < 0.001). Similarly, the aberrant expression rates of E‐cadherin and β‐catenin in bladder cancer tissues were significantly higher than those in normal bladder tissues (P < 0.001 for both comparisons). The expression of each molecule was correlated with tumor pathological grade and clinical stage (P < 0.05 for all comparisons), but not with tumor number and size (P > 0.05 for all comparisons). Furthermore, negative correlations were found between the expression intensities of p130Cas and E‐cadherin or β‐catenin in transitional cell carcinoma of the bladder (P < 0.05 for both comparisons). Conclusions: p130Cas, E‐cadherin and β‐catenin might represent useful predictors of malignant degree of transitional cell carcinoma of the bladder.     

Prognostic Significance of p53 Protein Accumulation in Stage pT1 Transitional Cell Carcinoma of the Bladder

Objective: Mutations in the tumour suppressor gene p53 results in the production of a mutant type, dysfunctional p53 protein which can readily be detected in the cell nucleus by immunohistochemical staining. This study aims to investigate the association of nuclear p53 protein accumulation with the clinical outcome of stage pT1 transitional cell carcinoma of the bladder which is renowned for high rates of recurrence and progression. Methods: TUR samples of the tumours from fifty-two patients with primary stage T1 bladder cancer were analyzed immunohistochemically using the standard avidin-biotin peroxidase method for nuclear p53 accumulation. Status of p53 immunostaining was correlated with tumour recurrence, disease progression and three-year survival of each patient. Results: The rate of tumour recurrence in pT1 bladder cancer was 36% in patients with tumours stained negatively for p53 protein and 78% in patients with tumours stained positively for p53 protein. Disease progression was seen in 15% of p53 (-) patients and in 56% of p53 (+) patients. Conclusions: In stage pT1 bladder tumours p53 nuclear accumulation indicates higher rates of tumour recurrence and disease progression. Accordingly, in patients who have pT1 bladder tumours with nuclear p53 accumulation, institution of more aggressive therapy should be considered and early radical therapeutic modalities should be offered to these patients.     

Clinical Significance of Nuclear p53 Protein Accumulation in Bladder Cancer

Objectives: To investigate the correlation of nuclear p53 accumulation with disease outcome in a cohort of patients with transitional cell carcinoma of the bladder. Methods: A total of 90 patients (11 female, 79 male) with transitional cell carcinoma of the bladder were included in this study. Tumour samples from the primary tumour were analysed by immunohistochemistry for nuclear accumulation of p53 protein. Outcome of each patient was recorded and investigated for a possible relation with p53 status. Results: Nuclear p53 deposition was determined in 22 specimens. The nuclear p53 deposition was seen in less than 20% of the nuclei examined in 13 and more than 20% in 9 cases. No stromal staining was observed. Nuclear p53 deposition was present in 15.2% (7/46) of grade 2 tumours, and 34% (15/44) of grade 3 tumours (p=0.037). Stage distribution revealed 15.5% (5/33) positivity in stage pTa, 25.8% (8/31) in pT1 and 34% (9/26) in stage pT2–3 tumours. Tumours with p53 nuclear accumulation had a higher rate of recurrence and progression and shorter survival. Conclusion: Results of the current study indicate p53 as an important factor in determination of biological behaviour of bladder cancer.     

Reciprocal expression of bcl-2 and p53 oncoproteins in urothelial dysplasia and carcinoma of the urinary bladder

In order to investigate if and when the bcl-2 oncoprotein is activated in bladder tumorigenesis and its relationship with p53 overexpression and patient survival, we studied bcl-2 and p53 expression immunohistochemically in matched normal urothelium, dysplasia and cancer specimens selected by step-sectioning from 54 radically resected bladders for non-metastatic transitional cell carcinoma (TCC). In normal urothelium and mild dysplasia, bcl-2 was restricted to the basal cell compartment, while in moderate and severe dysplasia its expression was detectable also in the upper regions. Excess bcl-2 immunoreactivity was found in 27 (50%) of carcinomas, and a larger proportion of high-grade TCCs showed bcl-2 expression compared with that of low-grade TCCs (P < 0.05). Overexpression of p53 protein showed a increasing trend toward the progression of bladder tumorigenesis (P < 0.01) and a significant reciprocal correlation was found between bcl-2 and p53 expression in either various dysplasias (P < 0.01) or carcinoma (P < 0.05). With the evolution from mild dysplasia to carcinoma in individual cases, loss of bcl-2 expression was more frequently observed in superficial (P < 0.02) or low-grade carcinoma (P < 0.05) than in muscle-invasive or high-grade carcinoma. Furthermore, patients with negative immunostaining for both bcl-2 and p53 in cancer lesions had a significantly more favorable prognosis compared with those with positive immunostaining for the oncoproteins (P < 0.05), although bcl-2 by itself did not predict patient survival. We suggest that aberrant activated bcl-2, which is seen earlier than p53, appears to facilitate bladder tumorigenesis and to enhance tumor aggression in some extent. Received: 22 October 1997 / Accepted: 2 January 1998     

Cell Cycle Proteins Predict Recurrence in Stage II and III Colon Cancer

Purpose

To investigate the prognostic value of multiple cell cycle-associated proteins in a large series of stage II and III colon cancers.

Methods

From formalin-fixed, paraffin-embedded tumor samples of 386 patients with stage II and III colon cancer, DNA was isolated and tissue microarrays were constructed. Tissue microarray slides were immunohistochemically stained for p21, p27, p53, epidermal growth factor receptor, Her2/Neu, β-catenin, cyclin D1, Ki-67, thymidylate synthase, and Aurora kinase A (AURKA). Polymerase chain reaction–based microsatellite instability analysis was performed to allow for stratification of protein expression by microsatellite instability status.

Results

Overall, low p21, high p53, low cyclin D1, and high AURKA expression were significantly associated with recurrence (P = 0.01, P < 0.01, P = 0.04, and P < 0.01, respectively). In stage II patients who did not receive adjuvant chemotherapy (n = 190), significantly more recurrences were observed in case of low-p21 and high-p53-expressing tumors (P < 0.01 and P = 0.03, respectively). In stage III patients who did not receive chemotherapy, high p53 expression was associated with recurrence (P = 0.02), and in patients who received chemotherapy, high AURKA expression was associated with relapse (P < 0.01). In patients with microsatellite stable tumors, high levels of p53 and AURKA were associated with recurrence (P = 0.01 and P < 0.01, respectively). Multivariate analysis showed p21 (odds ratio 1.6, 95% confidence interval 0.9–2.8) and AURKA (odds ratio 2.7, 95% confidence interval 1.3–5.6) to be independently associated with disease recurrence.

Conclusions

p21, p53, cyclin D1, and AURKA could possibly be used as prognostic markers to identify colon cancer patients with high risk of disease recurrence.

     

Expression Level of Valosin-Containing Protein (p97) Is Correlated With Progression and Prognosis of Non-Small-Cell Lung Carcinoma

Background: Valosin-containing protein (VCP; also known as p97) was shown to be associated with antiapoptosis and metastasis via activation of a nuclear factor-B signaling pathway. Our previous study showed that the VCP expression level correlated with the disease recurrence rate and prognosis of patients with hepatocellular carcinoma and gastric carcinoma. This study was designed to evaluate the prognostic significance of VCP expression in non-small-cell lung carcinoma (NSCLC).Methods: VCP expression in 207 patients with NSCLC (133 men and 74 women) aged 35 to 78 years (median, 62 years) was examined by immunohistochemistry, in which staining intensity in tumor cells was categorized as weaker than (level 1) or equal to or stronger than (level 2) that in endothelial cells.Results: Sixty-nine (33.8%) cases showed level 1 and 135 (66.2%) showed level 2 VCP expression. The frequency of the following was higher in patients with level 2 expression than in those with level 1 expression: male sex (P < .01), histological subtype of squamous cell carcinoma (P < .05), and smoking (P < .05). Patients with level 2 expression had poorer disease-free and overall survival rates (P < .05 and P < .01, respectively) than those with level 1 expression. Multivariate analysis revealed VCP expression and pathologic T (pT) and pN classifications to be independent prognostic factors for both disease-free and overall survival and showed vascular invasion to be an independent prognostic factor for overall survival. VCP level was a prognosticator for overall survival in both the early (pT1) and advanced (pT2–3) group of the pT classification (P < .05 for both).Conclusions: The prognostic significance of VCP expression in NSCLC was demonstrated.     

Correlation of p53 with the clinicopathologic features and prognosis of colorectal adenocarcinoma

Immunohistochemical staining of p53 was performed using an anti-p53 mouse monoclonal antibody, Pab1801, on 67 colorectal adenocarcinoma specimens to determine the prognostic value of p53 in colorectal cancer patients. Of a total of 67 tumors examined, p53 was detected in 34, but the rate of positive staining for p53 did not correlate with the clinical stage of disease. In 59 patients undergoing curative resection of the tumor, there was no significant difference in the recurrence rate (P = 0.137) or the disease-free survival rate between 28 patients with p53 positive tumors and 31 with p53 negative tumors (P = 0.135).     

h-prune Is an Independent Prognostic Marker for Survival in Esophageal Squamous Cell Carcinoma

Background  The human homologue of Drosophila prune (PRUNE, which encodes h-prune) protein interacts with glycogen synthase kinase 3 and promotes cell motility. The aim of our study was to investigate the impact of immunohistochemically detected h-prune expression on the survival of patients with esophageal squamous cell carcinoma (ESCC). Methods  Immunohistochemical staining of h-prune was performed for 205 surgically resected specimens of ESCC. Results  In total, 43 (21%) of 205 ESCC cases were positive for h-prune. h-prune-positive ESCC cases showed a more-advanced T stage (P < 0.0001), N stage (P < 0.0001), and tumor stage (P < 0.0001) than h-prune-negative ESCC cases. In the group of 116 stage II and III ESCC cases, recurrence of ESCC was frequently found in h-prune-positive cases. In patients with lung recurrence, the tumors were more likely to be h-prune positive than h-prune negative. Univariate analysis revealed that T stage (P < 0.0001), N stage (P < 0.0001), tumor stage (P < 0.0001), and h-prune staining (P < 0.0001) were significant prognostic factors for survival. Multivariate analysis indicated that N stage (P = 0.0182) and h-prune staining (P < 0.0001) were independent predictors for survival. Conclusions  These results indicate that immunostaining of h-prune is useful to identify patients at high risk for recurrence or poor prognosis associated with ESCC.     

Evaluation of p53 nuclear accumulation in low- and high-grade (WHO/ISUP classification) transitional papillary carcinomas of the bladder for tumor recurrence and progression

OBJECTIVES: To evaluate the association of p53 nuclear accumulation with recurrence and progression in transitional cell carcinomas of the bladder and to examine the distribution of p53 in low-grade and high-grade transitional cell carcinomas according to the World Health Organization/International Society of Urological Pathology classification. PATIENTS AND METHODS: Nuclear accumulations of p53 were examined in a total of 99 patients with transitional cell carcinoma between May 1995 and October 1999. The mean age was 64 years. There were 94 (95%) men and 5 (5%) women. Following resection, surgical specimens were examined, and p53 accumulation with a 20% cutoff value was accepted as positive staining. Of the 99 patients, 52 (53%) had histologically superficial bladder tumors, and 47 (47%) had invasive tumors. Data concerning grade, stage, number of recurrences, and disease progression were available for each patient. RESULTS: The median follow-up period was 55 months. 60 of the 99 patients (61%) had p53 overexpression. The difference for p53 overexpression between low-grade and high-grade tumors was significant (p < 0.05). In low- and high-grade tumors, there was no significant relationship for recurrence between p53-positive and p53-negative groups. But there was a statistically significant relationship between progression and histological grade of the tumors. p53 had no significant relationship with tumor recurrences (p > 0.05), but its relationship with progression was statistically significant (p < 0.05). CONCLUSIONS: We did not find a correlation between tumor recurrence and p53 overexpression, but p53 overexpression has a predictive value in determining tumor progression. High-grade tumors had higher p53-positive values than low-grade tumors. This group of patients should be considered for radical therapies on the basis of other prognostic parameters.     

Histopathological and biological prognostic risk factors in low stage testicular nonseminomatous germ cell tumors Implikationen für eine risikoadaptierte Therapie

Purpose of our study was to develop a reliable model to define clinical stage I nonseminomatous germ cell tumors (NSGCT) being at low risk and at high risk for occult retroperitoneal metastases based on pathohistological and immunohistochemical parameters in order to stratify the therapeutic approach. 3–5 paraffin-embedded formalin fixed tissue blocks of 149 clinical stage I NSGCT were available from all patients and were analyzed for histopathological features associated with pathological stage: presence/absence of vascular invasion, presence/absence of tunical invasion, percentage of each histological cell type present in the primary tumor. Immunohistochemical expression of MIB-1, p53, bcl-2, cathepsin D and e-cadherin was evaluated using a semiquantitative scoring ystem. Statistical analysis was performed by univariate and multivariate logistic regression models. Percentage of embryonal carcinoma [%EC (p < 0.001)] and presence of vascular invasion [VI (p < 0.0001)] were the most significant independent risk factors associated with pathological stage II disease. Combination of %EC and VI allowed correct prediction of final pathological stage in 88 % of patients. Cut-off values including both variables identified correct pathological stage in 131/149 patients (88 %). Less than 45 % EC and absence of VI correctly identified pathological stage I disease in 91.5 %; more than 80 % EC and presence of VI correctly predicted pathological stage II in 88 % of the patients. %EC and presence/absence of VI appear to be reliable prognosticators to identify both patients at high risk and at low risk for occult retroperitoneal disease. P53, bcl-2, MIB-1, cathepsin D and e-cadherin did not appear to be of prognostic value in clinical stage I NSGCT.     

Evaluation of p21WAF1/CIP1 and cyclin D1 expression in the progression of superficial bladder cancer

Immunoreactivity of p21^WAF1/CIP1 and cyclin D₁ proteins was assessed in a cohort of 207 patients with superficial (pTa-pT₁) bladder cancer followed up for a mean of 4.9 years. The results of the immunostainings were compared with T category, WHO grade, tumor cell proliferation rate (MIB-1 score), the expressions of p53 and bcl-2 as well as survival. Sixty-eight percent and 75% of the tumors were p21^WAF1/CIP1 positive (≥5% of cells positive) and cyclin D₁ positive (≥10% of cells positive), respectively. The p21^WAF1/CIP1 expression was related to cyclin D₁ immunolabelling (P < 0.001) but not to the other variables studied. The expression of cyclin D₁ was inversely associated with T category (P=0.001), WHO grade (P=0.006), MIB-1 score (P=0.014), p53 expression (P=0.001), and bcl-2 (P=0.011) immunoreactivity. In univariate analysis, T category (P=0.0001), WHO grade (P < 0.0001), MIB-1 score (P < 0.0001), bcl-2 (P=0.0092), p53 (P=0.0016) and p21^WAF1/CIP1 (P=0.009) expressions were significant prognostic factors with regard to tumor progression, whereas cyclin D₁ was without any prognostic significance (P=0.1). Out of 123 p21 positive tumors 21 progressed, whereas only 2 out of 58 p21 negative tumors progressed. In multivariate analysis, the MIB-1 score was the only independent predictor of cancer-specific survival (P=0.03), whereas tumor grade (P=0.002) and cyclin D₁ expression (P=0.04) were independent predictors of tumor recurrence. Only the WHO grade (P=0.04) retained its prognostic value indicating the risk of progression. We suggest that in superficial bladder cancer p21^WAF1/CIP1 and cyclin D₁ immunohistochemistry provide no additional prognostic information compared with already established prognostic factors for predicting the risk of progressive disease. Received: 13 September 1999 / 22 March 2000     

Expression of pS2 protein and its relation with the Ki-67 proliferative indices and tumor recurrence in superficial bladder carcinomas

OBJECTIVE: To investigate the expression and possible role of pS2 protein as a predictor of tumor recurrence in superficial transitional cell carcinoma of the bladder and to determine its relation with tumor stage, grade, size, number, recurrence and proliferative activity. METHODS: Paraffin sections of transurethral resection material from 80 patients with superficial transitional cell bladder carcinoma were stained with pS2 and Ki-67 antibodies using the standard streptavidin biotin immunoperoxidase method. Cytoplasmic pS2 staining was scored on a scale of 1-3 and the Ki-67-labelling index was determined as a percentage of positively staining tumor cells. RESULTS: An inverse relationship was found between pS2 expression and Ki-67 index (p<0.001). pS2 expression showed no relation with any clinicopathological prognostic parameters as well as the recurrence rate. The recurrence rate was only associated with increased tumor number (p = 0.05), while the time to first recurrence was significantly related to tumor size, proliferative activity and tumor grade (p = 0.04, p<0.001, and p = 0.03, respectively). On the other hand, higher tumor grade was correlated with increased tumor number, Ki-67 index and tumor stage (p = 0.016, p = 0.006, and p<0.001, respectively). CONCLUSION: pS2 expression is associated with a low proliferative potential of superficial transitional cell carcinoma of the bladder, while it does not seem to be related to the recurrence rate of the tumor and other prognostic factors. Tumor size and proliferative activity may aid in the estimation of the time to the first recurrence.     

Impact of the EpCAM expression on biochemical recurrence-free survival in clinically localized prostate cancer

BackgroundThe epithelial cell adhesion molecule (EpCAM) is a transmembrane glycoprotein that was originally identified as a marker for carcinoma, attributable to its high expression on rapidly proliferating tumors of epithelial origin. The role of EpCAM is not limited to cell adhesion but includes diverse processes such as signaling, cell migration, proliferation, and differentiation.ObjectiveSeveral studies investigated EpCAM expression in prostate carcinoma but none of them confirmed its prognostic role. The aim of our study was to investigate EpCAM expression and its relationship with established prognostic features in prostate carcinoma.Materials and methodsThe study included a cohort of 102 patients treated with radical prostatectomy for clinically localized prostate carcinoma. Immunohistochemistry was performed to evaluate the EpCAM expression in prostate cancer and non-neoplastic prostate tissue. The percentage of positively stained carcinoma and benign glands was examined in the whole mount of the chosen slide.ResultsThe extent of EpCAM expression was significantly higher in malignant than in benign prostatic tissue (P < 0.001). EpCAM expression in prostate cancer was associated with established features indicative of worse prognosis, such as preoperative (P = 0.009) and postoperative (P = 0.004) Gleason score and follow-up time (P < 0.001). Patients with higher preoperative and postoperative Gleason score and short follow-up time had tumors with a significantly higher expression of EpCAM. Negative correlation of follow-up time and EpCAM expression indicated that tumors in patients with biochemical recurrence (BCR) harbored higher EpCAM expression. Moreover, expression of EpCAM was significantly higher in patients with BCR compared with patients without BCR (P < 0.001). Tumors in T3 stage of the disease showed significantly higher EpCAM expression compared with T2 tumors (P = 0.002). Univariate (P < 0.001) and multivariate (P < 0.001) analyses showed that EpCAM expression was a significant predictor of shorter biochemical recurrence free-survival.ConclusionOur results confirmed high level of EpCAM expression in prostate cancer and support its potential role in prostatic cancer progression. In addition, EpCAM could serve as an additional prognostic marker for the recognition of patients with an increased risk of disease recurrence that need introduction of secondary therapy.     

Expression of cyclooxygenase-2 in normal urothelium, and superficial and advanced transitional cell carcinoma of bladder

PURPOSE: We compared the differential expression of cyclooxygenase-2 in normal bladder tissue, primary bladder transitional cell carcinoma and transitional cell carcinoma metastases to lymph nodes, and determined whether cyclooxygenase-2 expression is associated with molecular alterations commonly found in bladder transitional cell carcinoma and clinical outcomes after radical cystectomy. MATERIALS AND METHODS: Immunohistochemical staining for cyclooxygenase-2, survivin (Novus Biologicals, Littleton, Colorado), p21, p27, pRB, p53, MIB-1, Bax, Bcl-2, cyclin D(1) (Dakotrade mark), cyclin E (Oncogene, Cambridge, Massachusetts) and caspase-3 (Cell Signaling, Beverley, Massachusetts) was performed on archival bladder specimens from 9 subjects who underwent cystectomy for benign causes, 21 patients who underwent transurethral resection and 157 consecutive patients after radical cystectomy, and on 41 positive lymph nodes. RESULTS: Cyclooxygenase-2 was expressed in none of the 9 normal bladder specimens (0%), 52% of transurethral resection specimens, 62% of cystectomy specimens and 80% of lymph nodes involved with transitional cell carcinoma. Cyclooxygenase-2 expression was associated with higher pathological stage, lymphovascular invasion and metastases to lymph nodes (p=0.001, 0.045 and 0.002, respectively). Cyclooxygenase-2 expression was associated with altered expression of p53 (p=0.039), pRB (p=0.025), cyclin D1 (p=0.034) and caspase-3 (p=0.014). On univariate analysis cyclooxygenase-2 expression was associated with an increased risk of disease recurrence and bladder cancer specific mortality (p=0.0189 and 0.0472, respectively). However, on multivariate analysis only pathological stage and metastases to lymph nodes were associated with disease recurrence (p<0.001 and <0.001) and survival (p<0.001 and 0.015, respectively). CONCLUSIONS: Cyclooxygenase-2 is not expressed in normal bladder urothelium. Cyclooxygenase-2 over expression is associated with pathological and molecular features of biologically aggressive disease, suggesting a role for cyclooxygenase-2 in bladder cancer development and invasion.     

p53 Gene mutations in superficial bladder cancer

OBJECTIVES: To assess the presence of p53 gene mutations in superficial tumors of the urinary bladder (transitional cell carcinoma) and their relationship to classic prognostic factors for cancer recurrence and progression. To analyze the implication of these mutations on the P53 protein structure. MATERIALS AND METHODS: Observational, cross-sectional study of 90 consecutive patients, 60 with superficial transitional cell carcinoma (pTa and pT1) and 30 without neoplastic disease (control group). Samples of bladder tumor and control normal mucosa were analyzed by polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) to detect p53 mutations in exons 5-9. Automatic sequencing was used to characterize the mutations and their effect on the P53 protein was analyzed. Bivariate analysis was used to assess the association with other prognostic factors. RESULTS: PCR-SSCP found no mutations in any control group patient, whereas 38.3% of patients with superficial transitional cell carcinoma had one or more mutations in the exons analyzed. Thirty mutations were sequenced; all were point mutations and 86.67% were considered relevant for the P53 structure. A total of 93.3% of the mutations were located in highly conserved regions and 73.3% in mutational hot spots. The highest cell differentiation grades and pT1 stage were associated with a higher incidence of p53 gene mutations. Previous recurrences and other tumor-related histological variables were not associated with a higher percentage of mutations. CONCLUSION: Mutations at p53 did not appear in healthy bladder mucosa and were significantly more frequent in pT1 and high-grade (G-II and G-III) tumors. All mutations detected were point mutations and most caused considerable P53 structural abnormalities, implying major repercussions on P53 function. These data suggest that certain p53 mutations may have prognostic value, even though they were not associated with other classic recurrence and tumor progression parameters. Future analyses of the progress of patients with superficial bladder transitional cell carcinoma and mutated p53 will help clarify this aspect.     

Comparative Study of the Expression of p53, Ki67, E-cadherin and MMP-1 in Verrucous Hyperplasia and Verrucous Carcinoma of the Oral Cavity

Context Oral verrucous carcinoma (OVC) and oral verrucous hyperplasia (OVH) may be clinically and histologically similar. Problems separating these lesions are compounded by poorly oriented tissue sections and biopsies failing to demonstrate lesional margins. Objective To distinguish OVC from OVH utilizing an immunohistochemical panel (p53, matrix metalloproteinase-1, E-cadherin, Ki67) shown to be useful in differentiating pseudoepitheliomatous hyperplasia from oral squamous cell carcinoma of the head and neck. Materials Twenty-eight cases of OVH and thirty-two cases of OVC studied. Diagnoses were confirmed by two pathologists. Formalin-fixed, paraffin-embedded archival material was used for immunohistochemistry (avidin–biotin immunoperoxidase technique). Results More diffuse nuclear staining of p53 and Ki67 was detected in the OVC cases compared to the OVH cases (P < 0.001). There was statistically significant increased staining within adjacent stromal cells for matrix metalloproteinase-1 (P < .05) in the OVC cases. E-cadherin demonstrated diffuse membranous staining in both groups. Conclusion Ki67, p53, and MMP-1 demonstrated significant staining trends. Although a properly oriented hematoxylin–eosin-stained section including normal marginal tissue is considered to be the gold standard for differentiation of OVH and OVC, this immunohistochemistry panel may serve as a useful diagnostic adjunct in difficult cases.     

p53 Accumulation Is a Prognostic Factor in Intestinal-Type Gastric Carcinoma but Not in the Diffuse Type

Background: The prognostic value of p53 nuclear accumulation in gastric cancer is still unclear, as shown by the discordant results still reported in the literature. In this study, we evaluated the correlation between p53 accumulation and long-term survival of patients resected for intestinal and diffuse-type gastric cancer.Methods: Eighty-three patients with carcinoma of the intestinal type and 53 patients with carcinoma of the diffuse type were included in the study. Immunohistochemical staining of the paraffin sections was performed by using monoclonal antibody DO1; cases were considered positive when nuclear immunostaining was observed in 10% or more of the tumor cells. Prognostic significance of different variables was investigated by univariate and multivariate analysis.Results: p53 positivity was found in 51.8% of intestinal-type and 50.9% of diffuse-type cases. No significant correlation between the rate of p53 overexpression and age, sex, tumor location, tumor size, depth of invasion, lymph node involvement, distant metastases, and surgical radicality was found in the two groups of patients. A statistically significant difference in survival rate was observed between p53-negative and p53-positive cases in the intestinal type (P < .05), confirmed by multivariate analysis (P < .005; relative risk = 3.09). On the contrary, no correlation with survival was found in diffuse-type cases according to p53 overexpression.Conclusions: These results suggest that the immunohistochemical detection of p53 accumulation is a useful indicator of poor prognosis in the intestinal but not in the diffuse type of gastric cancer, and are indicative of distinct molecular pathways and pattern of progression in the two histotypes.