FGFR3, P53and MCM2Protein Expression In Urothelial Carcinoma Of The Bladder And Their Relationship To Prognostic Variables

Objective:To summarize the clinicopathological features of patients with urothelial carcinoma of the bladder, and analyze the association of established clinical and pathological characteristics with tumor recurrence and progression; To evaluate the expression and prognostic value of FGFR3, p53and MCM2protein expression in patients with urothelial bladder cancer and to investigate their correlation with eatablished prognostic factors; To provide theoretical basis for selecting the appropriate type and timing of therapy and assessing the prognosis.Materials and methods:Part I:The formalin-fixed paraffin-embedded samples of658cases of urothelial carcinoma of the bladder were collected in the Department of Pathology, Fudan University Shanghai Cancer Center. The cases were diagnosed between2006and2010. The clinical information and pathological data of the patients were acquired from the database of the hospital. All histopathologic materials from the diagnostic tumors were reviewed by two senior pathologists. Staging and grading of each tumor was done according to the TNM classification and the2004WHO classification criteria for malignant tumors of the urinary tract, respectively, and assess the tumor growth pattern. The relationship between clinicopathological parameters and recurrence and progression in UCB was investigated by univariate and multivariate analysis.Part II:317cases were randomly selected from the above658cases. A tissue microarray (TMA) of166superficial urothelial carcinoma (Ta-T1) and151invasive urothelial carcinoma (T2-T4) and20nonneoplastic tissue controls was constructed. Immunohistochemistry(IHC) was used to detect the expression of FGFR3、p53and MCM2. Additional covariates measured included patient age, gender, pathological stage, histologic grade, tumor size, number, papillary growth pattern and type of surgery,et al. The relationship of these biomarkers and the association between biomarkers and clinicopathologic parameters and recurrence and prognosis was analyzed. Results:Part Ⅰ:(1)The clinicopathologic features:The median follow-up period was22.3months.The mean patient age was55years old ranging from20to90years. Most patients were between50and70years of age. Male to female ratio was about5:1. The most frequent complaint was gross hematuria in76%of the patients. At initial diagnosis, about50%patients had stage Ta tumors and29%had stage T1tumors.12.7%patients had muscle invasive urothelial carcinomas (pT2-pT4). The percentage of low-grade tumors and tumors with papillary growth pattern was49.9%and69.6%, respectively. The recurrence and progression rate were35.6%and5.9%, respectively.(2)The relationship between clinicopathologic parameters:97.2%invasiveUCB and41.3%superficial UCB were of high grade, it had a statistically significant (p<0.001).92.4%low-grade UCB and45.6%high-grade UCB,78.8%superficial UCB and17.0%invasive UCB had papillary growth pattern (both p<0.001).(3)The relationship between clinicopathologic parameters and recurrence and progression:The multivariate analysis showed that pathological stage was associated with recurrence (p<0.05).High-grade UCB tend to recur more frequently than low-grade UCB, but the statistical analysis was not significant (p=0.085).In univariate analysis, gender (p=0.035),pathological grade (p<0.001), tumor growth pattern (p<0.001) and pathological stage (p<0.001) were associated with tumor progression (p<0.05).Only pathological grade(p=0.028),pathological stage(p=0.043) and tumor growth pattern(p=0.022) were independently associated with tumor progression on multivariate analysis.Part Ⅱ:(1) The expression of FGFR3, p53and MCM2protein:Well-differentiated UCB and superficial UCB (pTa and pT1) tend to have more frequently cytoplasmic FGFR3expression than poorly-differentiated UCB and invasive UCB (pT2to pT4)(all p<0.001). However, poorly-differentiated tumors (p=0.003) and invasive tumors (p<0.001) tend to express more frequently nuclear FGFR3than well-differentiated tumors and superficial tumors. UCB with papillary growth pattern tend to have more cytoplasmic FGFR3expression (p<0.001) and less nuclear FGFR3expression(p<0.001) than UCB with nonpapillary growth pattern. Low-grade UCB had low expression level of p53than high-grade UCB (p<0.001), superficial UCB had low expression level of p53than invasive UCB (p=0.024). The expression level of p53 was lower in tumors with papillary growth pattern (p<0.001). High grade tumors displayed a higher expression level than the remaining cases (p<0.001). Although invasive UCB displayed a higher expression level of MCM2protein, the correlation between MCM2and pathological stage was not statistically significant (p=0.103). There was a significant association between MCM2and papillary status (p<0.001), that is UCB with papillary growth pattern tend to have a lower MCM2expression level, and MCM2preferentially stained the basal cell layers in tumors with papillary growth pattern.(2) The relationship between FGFR3,p53and MCM2protein:A statistically significant strong correlation was established between p53and FGFR3-C (Spearman p=-0.25,/?<0.001),MCM2and FGFR3-C(Spearmanp=-0.277,p<0.001),MCM2and FGFR3-N(Spearmanp=0.271,p<0.001),p53and MCM2(Spearmanp=0.343,p<0.001). Tumors that displayed a higher expression level of p53protein tend to have a more frequent nuclear FGFR3expression, however, this parameter was not statistically significant (p=0.076).(3) Factors in relation to tumor recurrence:In univariate analysis, tumor number (solitary vs multiple,p=0.03) and gender (p=0.031) were associated with tumor recurrence. In multivariate analysis, gender (p=0.037), tumor size (p=0.033) and number (p=0.015), pathological grade (p=0.006) and p53protein expression (p=0.017were associated with recurrence.(4) Recurrence-free survival analysis:High-level MCM2expression was significantly associated with shorter RFS when using a cutoff of10%(p=0.006), while the median value35%did not reach a statistical significance (p=0.383). Tumors with positive cytoplasmic FGFR3expression (moderate/strong) tend to have longer RFS than tumors with negative/weak FGFR3expression, although the statistical analysis was not significant (p=0.156). Patients with positive nuclear FGFR3tend to have longer RFS than patients with FGFR3negativity in pT2-pT4UCB, however, the parameter was not statistically significant (p=0.108). Patients without FGFR3expression (both cytoplasmic and nuclear negative) had shorter RFS than patients with cytoplasmic or/and nuclear FGFR3expression (p=0.020). Multivariate Cox regression analysis demonstrated that only tumor grade was independently associated with tumor recurrence-free survival (p=0.044).(5) Disease-free survival analysis:Kaplan-Meier curves using the log-rank tests showed men (p=0.004), low grade tumors (p<0.001), papillary growth pattern (p<0.001) and stage Ta/T1(p=0.002) had longer RFS. Patients without FGFR3expression had shorter DFS than patients with cytoplasmic or/and nuclear FGFR3expression (p=0.098). While using10%as a cutoff value, high-level MCM2expression was significantly associated with shorter RFS (p=0.011). When using the median value35%as a cutoff, high-level MCM2expression was significantly associated with worse survival in stage Ta/T1tumors (p=0.009),but not in stage T2to T4tumors (p=0.585). p53protein expression scored as "++" was associated with better survival(p=0.022). Univariate Cox regression analysis showed that gender, pathological grade and stage, tumor growth pattern, MCM2expression (10%cutoff), were the parameters with significant predictive value. Multivariate Cox regression revealed that gender, tumor number and pathological grade have an independent prognostic value in predicting survival.Conclusions:(1)Majority of the UCB in Fudan University Shanghai Cancer Center were stage pTa-pT1, most of the low-grade UCB and superficial UCB had papillary growth pattern. UCB had a high incidence of recurrence.(2)Only pathological stage was an independent predictor of recurrence. Tumor grade, pathological stage and tumor growth pattern were independently associated with tumor progression.(3)Superficial, low-grade and papillary growth UCB had higher FGFR3cytoplasmic expression than the remaining.However, invasive, high-grade and nonpapillary growth UCB had higher FGFR3nuclear expression.(4)Superficial, low-grade and papillary growth UCB had lower expression level of p53than invasive, high-grade and nonpapillary growth UCB.(5)Invasive, high-grade and nonpapillary growth UCB had higher expression level of MCM2than the remaining. When using10%as a cutoff value, patients with MCM2expression had shorter RFS and DFS.(6)The expression of FGFR3, p53and MCM2protein had a significant association with pathological grade, stage and growth pattern. There was an association between each other of FGFR3, p53and MCM2protein.(7)Gender, tumor size and number, pathological grade and p53protein expression were predictors of recurrence.