The total number of apoptotic cells in 10 randomly selected fields was counted. The apoptotic index (AI) was calculated as the percentage of positive staining cells, namely AI = number of apoptotic cells × 100/total number of nucleated cells. Statistics Results were expressed as mean ± standard deviation (SD) and were analyzed with a one-way ANOVA and SPSS18.0 software package used to perform statistical analysis. A value of P < 0.05 was considered significant between the experimental buy Ispinesib groups compared with other groups. Results Expression of ATM in ATM AS-ODNs transfected Hep-2 cells To analyze the expression of ATM in mRNA and protein level in Hep-2 cells, real-time fluorescent quantitative PCR and western blot assay were

used respectively. It is evident that there were no significant differences among the groups treated with buy SGC-CBP30 liposome alone, with Sen-ODNs and with Mis-ODNs after 72 hours treatment (P > 0.05; Figure 1). However when incubated with liposome formulations of ATM AS-ODNs, the relative ATM mRNA expression was only about 11.03 ± 2.51% to the untreated Hep-2 cells, which showed a significantly reduced expression of ATM

mRNA (P < 0.05;Figure 1). As shown in Figure 2, ATM protein expression was learn more also significantly reduced by ATM AS-ODNs compared with Sen-ODNs and Mis-ODNs after 72 hours treatment (Figure 2A). The relative ATM protein expression of Hep-2 cells treated with ATM AS-ODNs was only about 48.14 ± 5.53% to the untreated cells (P < 0.05; Figure 2B). But there was no significant difference among the group treated

with liposome alone, the group treated with Sen-ODNs, the group treated with Mis-ODNs and the group of control untreated Hep-2 cells (P > 0.05; Figure 2B). Figure 1 Real-time quantitative PCR analysis of ATM mRNA expression. Liposome formulations of ATM AS-ODNs decreased expression of ATM mRNA was notably lower than that of other groups. There are no significant differences among liposome-treated group (Lipo), Sen-ODNs (Sen-Lipo) treated group and Mis-ODNs (Mis-Lipo) treated group (P > 0.05). P < 0.05, AS-ODNs (AS-Lipo) treated group compared with other groups. Figure 2 A Effect of ATM AS-ODNs on the expression of ATM protein in vitro. (A) Liposome formulations of ATM AS-ODNs dramatically reduced the expression of ATM protein compared with other groups. (B) There are no significant Thiamet G differences among liposome-treated group (Lipo), Sen-ODNs (Sen-Lipo) treated group and Mis-ODNs (Mis-Lipo) treated group (P > 0.05), while the group treated with ATM AS-ODNs notably different compared with other groups (**P <0.05). ATM AS-ODNs on clonogenic survival ability of Hep-2 cells after irradiation Cloning efficiency, P <0.05, was declined in cells transfected with ATM AS-ODNs compared with untreated cells or cells treated with control at the identical radioactive dose (Figure 3). After 4 Gy radiation, the survival fraction (SF4) revealed the cellular radio-induced apoptosis.