DSS was defined as survival without death due to ovarian cancer, and OS was defined as survival without

death due to any cause. Relapse was defined as symptomatic disease based on physical examination, imaging studies, and CA125 levels, or for patients initially diagnosed with benign disease, the subsequent development of malignant disease. On the basis of DSS, patients were divided into two relapse groups: recurrent disease (present) and no recurrent disease (absent). On the basis of CA125 level, patients were grouped as low (< 35 IU/ml) and high (≥ 35 IU/ml). For statistical analysis, medians of TS means were used. In all the subsequent analyses, the R Statistical Language [17] was used. All correlation coefficients presented in this manuscript are “rho” coefficients from Spearman rank test. Survival analyses, survival plots, and Cox proportional hazards regression models were generated by the package “survival” [18] and [19]. RG7420 in vivo The P values presented in the plots are derived from the log-rank test. The survival function

of Everolimus DSS and OS was estimated using the Kaplan-Meier method. To determine the effect on likelihood of survival of combinations of proteins/clinicopathologic variables, the tree-structured survival analysis was used [20]. Patient clinical and pathologic data are summarized in Table 1. The follow-up for both groups was 60 months. Benign and metastatic ovarian tumors were both of serous type to exclude potential variations in protein expression between tumor subtypes. The interobserver variation was 0.8 (for all assessed proteins in all cell types). The expression patterns of all proteins were initially characterized in EOC cells. Representative EOC staining patterns for each protein are illustrated and discussed in Figure 1. Relatively high expression (median TS > 3.5) was observed for all proteins except MMP2 (Figure 2). The TS for MMP2 was 1 indicating

minimal expression (data not shown), and thus, this protein EGFR inhibiton was not included for further analysis. EOC cell TS of expression of each expressed protein in all individual patients studied and overall median TSs for each protein are shown in Figure 2. Next, we assessed expression of protein targets in the endothelium and mesothelium of both groups. Representative images, together with a description of the staining patterns, are presented in Figure 3. Endothelial and mesothelial cell TSs of expression of each protein in all individual patients studied and overall median TSs for each protein are shown in Figure 2. The malignant group mesothelium expressed the highest levels of MMP9, VEGFA, and CL, while the endothelium was particularly immunoreactive for VEGFA and CL. Mesothelial and endothelial MMP2 immunoreactivity was mainly negative or weakly positive in both groups. MMP9 immunoreactivity exhibited mainly diffuse, cytoplasmic staining, with stronger perinuclear pattern of staining observed in the mesothelium.