The hierarchical clustering was carried out using the Cluster program (Pearson correlation, average linkage) and visualized by TreeView (University of California at Berkeley, CA) [20]. The functional profiling of each protein cluster was performed using the g:Profiler annotation tool (University of Tartu, Estonia) under the criteria of P < .05. We started by analyzing the effects of different PTEN mutations on disease-free survival (DFS) of patients BMN-673 with GBM, which reflects the effectiveness of treatment and the tendency for cancer recurrence. A total number of 586 patients with complete genomic sequencing and clinical data
from the TCGA GBM data set [16] were selected for analysis in this study. The spectrum of PTEN mutation in the TCGA GBM data set was similar to that reported previously [14], including missense (51.2%), nonsense (16.9%), frameshift (24.9), and other types of mutations (7%; Figure 1A). Using Cox propor- tional hazards analysis, we analyzed the independent effects of PTEN mutation, promoter methylation and expression (protein and mRNA levels by arrays), genomic instability, and Karnofsky performance scale on DFS of patients with GBM. Intriguingly, nonsense muta- tions of PTEN associated with significantly shorter DFS (median, 3.8 months) than other mutations or wild-type genotype (median, 7.2 months), displaying higher
HR of 3.26 (95% CI = 1.48-7.20; Figure 1, B and C ). On the contrary, missense or frameshift muta- tions showed no significant association with DFS of patients with GBM. Moreover, overexpression of PTEN protein also
associated with shorter www.selleckchem.com/products/MDV3100.html DFS (median, 6.0 months) than other cases (median, 7.0 months), with increased HR of 1.31 (95% CI = 1.07-1.61; Figure 1, B and D). No correlation was found between patient DFS and PTEN mRNA level or promoter methylation, the number of mutations (as revealed by genomic sequencing), or fraction of genome with copy number alteration (CNA, an indication of genomic instability) DNA Damage inhibitor in GBM cases. The different effects of PTEN mutations on DFS suggest that these mutations also confer distinct biologic consequences. Because loss of PTEN function has been linked to genomic instability and impaired DNA repair ability [21], we compared the number of mutations and fraction of genome with CNA in patients with GBM carrying different types of PTEN mutations. Although missense, non- sense, and frameshift mutations were all found to increase the fraction of genome with CNA (Mann-Whitney test, P < .05; Figure 2A), only nonsense mutation of PTEN associated with a higher number of mutations in GBM tissues (Mann-Whitney test, P < .05; Figure 2B). Furthermore, nonsense mutation of PTEN also linked to decreased levels of p53 and Gata3 proteins (Mann-Whitney test, P < .05), but such link was not evident for missense or frameshift mutations ( Figure 2, D and E).