Survival functions were compared using the log-rank (Mantel-Cox) test. higher Fuhrman grades also had higher risks for cancer-specific mortality (hazard ratio, 11.4) and Polyphyllin VII tumor progression (hazard ratio, 4.5). In summary, our study provides evidence of dysregulation of the mammalian target of rapamycin and hypoxia-induced pathways in papillary renal cell carcinoma. Immunohistochemistry for members of the mammalian target of rapamycin pathway and for HIF-1lacked prognostic significance in our cohort. was defined as the presence of pelvic recurrence or metastasis to distant sites. Overall mortality refers to all-cause death. 2.1. Immunohistochemistry Immunohistochemistry was performed for the following proteins: PTEN, phos-AKT, phos-S6, 4EBP1, c-MYC, p27, and HIF-1score was calculated for each tissue microarray spot as the sum of the products of the intensity (0 for unfavorable, 1 for weakly positive, 2 for moderately positive, and 3 for strongly positive) by the extent of immunoexpression (0C100%). The overall score used for subsequent statistical analysis was the pooled mean from the 4 spots of normal kidney and of tumor tissue. 2.3. Statistical analysis Because of the non-gaussian distribution of the population (DAgostino-Royston assessments for normality, .001), nonparametric tests were used in all instances. Scores of paired normal kidney and tumor were compared using the Wilcoxon matched-pairs signed rank test. Differences in scores of tumor Polyphyllin VII tissue were stratified by clinicopathologic variables and compared using the Mann-Whitney test or the Fisher exact test. Unadjusted and adjusted Cox models were built Polyphyllin VII to evaluate the hazard ratio of clinicopathologic variables and biomarkers expression as prognosticators of outcome. Cumulative hazards were plotted using Nelson-Aalen estimates. Survival functions were compared using the log-rank (Mantel-Cox) test. For statistical significance, a 2-tailed .05 was required for bivariate analysis. The threshold was adjusted using the ?idk correction when more than 1 test was applied to the same set of data. Hazard ratios were interpreted using 95% confidence intervals. Data were analyzed using STATA release 11 (StataCorp Inc, College Station, TX). 3. Results Clinicopathologic features of all patients are shown in Table 2. In papillary RCC, overall mortality and cancer-specific mortality were 24% and 11%, respectively. Tumor progression was observed in 17% of the patients. Table 2 Demographic and clinicopathologic features of 54 patients with papillary RCC treated by nephrectomy and phos-S6, respectively. Scores of phos-S6, 4EBP1, and HIF-1were higher in tumor than in normal kidney. Conversely, scores of p27 were lower in tumor than Polyphyllin VII in normal kidney. Finally, scores of c-MYC and phos-AKT were comparable in tumor and in normal kidney. These patterns of expression are shown in Fig. 2. Open in a separate windows Fig. 1 Immunohistochemistry for members of the mTOR pathway and related proteins in papillary RCCs. A, Loss of PTEN immunoexpression. B, Weak and sparse nuclear phos-AKT positivity. C, Phos-S6 cytoplasmic positivity. D, 4EBP1 cytoplasmic positivity. E, Unfavorable c-MYC expression. F, Nuclear and cytoplasmic p27 positivity. Open in a separate windows Fig. 2 Patterns of immunohistochemical expression of members of the mTOR and hypoxia-induced pathways in papillary RCCs. Scale axis corresponds to scores. Light gray box plots correspond to normal kidney. Dark gray box plots correspond to tumor tissue. Outliers are represented by hollow triangles (for normal kidney) and hollow Rabbit monoclonal to IgG (H+L)(HRPO) diamonds (for tumor tissue). The box plot for c-MYC expression was omitted because of the small number of patients with H scores higher than 0. Table Polyphyllin VII 3 Comparison of score means and 95% CI in papillary RCC and paired normal kidney score. 3.2. Predictors of outcome in papillary RCC On bivariate analysis of pathologic features and biomarkers, only HIF-1was associated with tumor size (Table 4). The same pattern was observed for HIF-1and T stage. None of the other biomarkers were associated with T stage, Fuhrman.