Prevalences and confidence intervals of single studies were evaluated using Clopper and Pearson method [21]. Correlation of the presence of the H1047R mutation with clinical-pathological features, p-values and confidence intervals were evaluated by means of logistic regression analysis. Correlation with survival was evaluated by means of log-rank test. For Cox multivariate regression, we selected the most informative variables among the models that included mutational status, using a ‘forward’ stepwise method. A p-value less than 0.05 was considered significant. For all the calculations and illustrations the R statistical software package
was used [22]. Results We analysed the sequences of exons 9 and 20 of the PIK3CA gene in 264 advanced gastric cancers. The list and frequency of mutations found are detailed in Table 2. A total of 42 cases (15.9%; 95% CI 11.7% – 20.9%) harbored at least one mutation Nutlin-3a supplier in the regions analyzed. All the mutations found were heterozygous this website missense single base substitutions. The most common mutation was H1047R selleck compound occurring at the active site of the kinasic domain in
exon 20 and representing 62% of all the mutations. The second most common mutation was Q546K that involves an aminoacid change in the helicase domain in exon 9 and represents 9.5% of all the mutations found. Table 2 Frequency of PI3KCA mutations found in 264 gastric cancers, by mutation type. Mutation Overall frequency (MSI only) Percent/total cases Percent/mutated cases Exon 9 E542K 2 0.76% 4.76% E545K 2 0.76% 4.76% Q546K 4 1.52% 9.52% Total Mutations (ex. 9) 8 3.03% Exon 20 M1043V 1 0.38% 2.38% H1047R 26 (8) 9.85% 61.90% H1048T 1 0.38% 2.38% Thiamine-diphosphate kinase G1050D 2 0.76% 4.76% T1052I 1 0.38% 2.38% T1053I 1 0.38% 2.38% D1056N 2 0.76% 4.76% L1067F 1 0.38% 2.38% Total Mutations (ex.20) 35 13.26% Total Mutations 42 15.91% We found two missense mutations namely T1052I and T1053I that were never reported before. The mutations were confirmed using a second pair of primers (see Additional File 1). Both mutations involve an aminoacidic change from threonin to isoleucin that implies a change
in the hydrophobic properties of the residues and may potentially affect the protein function. One case harboured two mutations namely E545K and L1067F, in exons 9 and 20, respectively. In our series, MSI cases only harbored the H1047R mutation. H1047R was, in fact, observed in 8 of 39 MSI cases and was significantly associated with MSI status (OR 3.0; 95% CI 1.0 – 7.9; Fisher’s test P = 0.035). The presence of mutation H1047R did not correlate with either survival or other clinical pathological features generally associated with MSI, possibly due to the small number of cases harboring the mutation. Furthermore, we did not observe any significant association between the presence of mutation and survival when considering MSI cases only.