In the MRC AML 15 trial, AML patients 60 years with favourable risk disease who

After transfer, samples were blocked with 5% MP PBST for 1 h and probed with antibodies against AP24534 価格 Sirt1, cleaved PARP, pospho H2AX pSer139 and beta Actin diluted in 5 MP PBST and incubated at 4 C overnight. The appropriate secondary antibody was applied at room temperature for 1 hr. Visualization was performed by enhanced chemilu minescence, Western blots signals were quantified using the ImageJ 1. 32 software after scanning of the films. Statistical analysis For correlation analysis of Sirt1 expression with clinic pathological parameters, the Fishers exact test or χ2 test for trends was applied. For univariate analysis we used the Kaplan Meier method and a Log rank test to probe for significance. For multivariate survival analysis the Cox proportional hazard method was used.

Variables found in univariate analysis to be significantly related to survival were included in the AT7519 溶解度 Cox models. For statistical analysis of cell cycle and MTT data, a two tailed t test was applied. For all statistical tests and methods, p values of 0. 05 were considered statistically significant. Statistical analyses were carried out with SPSS 15. 0 and Graph Pad Prism 4. Results Patients and tumor characeristics The patients demographics are listed in Table 1. The mean follow up time was 22. 1 months. During the study period, 89 patients died. The median survival was 13. 4 months and the median time to death was 10. 3 months, 65 patients were below the age of 65 and 64 patients above the age of 65, 118 PDAC were located in the head of the pancreas and 11 in the pan creatic corpus or tail.

Sirt1 expression in PDACs The specificity of the antibody used for immunohisto chemistry was corroborated by siRNA mediated knock down of Sirt1 in MiaPaCa 2 and PANC 1 cells and subsequent immunoblotting with the Sirt1 antibody. The knock down led to complete abrogation of the immunosignal as shown in buy Alisertib Figure 1. As exemplified in Figure 2, we observed a nuclear localization of Sirt1 in PDAC with a low expression in 72. 1% and a high expression in 27. 9% of the cases, respectively. Sirt1 was expressed by tumor cells with varying degrees of nuclear atypia, forming either neoplastic duct like structures, solid masses or single cell infiltrates within desmoplastic stroma.

When analyzed with regard to the morphological fea tures and tumor extent, the expression of Sirt1 was sig nificantly correlated to poor histological differentiation, There was no statistical difference in Sirt1 expression between early stage and advanced stage tu mors, Univariate survival analysis By univariate survival analysis, patients outcome was correlated with both tumor TNM and WHO stage, A borderline significance was observed for histological grade, The Kaplan Meier analysis of grouped Sirt1 expression was highly prognostic of poor overall survival for those patients with high Sirt1 expression with a mean postsurgical survival of 13. 0 vs. 54. 1 months, Multivariate survival analysis In multivariate Cox regression analysis, high Sirt1 expression was significantly related to shorter over all survival, in dependently of the degree of histological differentiation and WHO stage.