Study Of Akt Inhibitor In Combination With Rituximab On Treatment Of Burkitt Lymphoma

Objetive To investigate the effect of Akt inhibitor (AZD5363) in combination with rituximab on cell proliferation and apoptosis in Burkitt Lymphoma in vitro, and to explore its mechanism, which provided a theoretical basis for Akt inhibitor in combination with rituximab on treatment of Burkitt’s Lymphoma.Methods 1. Daudi cells were cultured in vitro, AZD5363 and rituximab were needed in various concentrations, CCK8 colorimetry was used to measure the proliferation of Daudi cells after 24 hours.2. Selected the concentration of AZD5363 and rituximab at 40 μmol/L and 50μg/mL for further study, and divided into 4 groups:control group, AZD5363 group, rituximab group and combined treatment group (AZD5363 plus rituximab), growth inhibition was detected by CCK8 colorimetry after 24,48,72 hours respectively; and cells apoptosis was detected by flow cytometry after 48,72 hours respectively.3. Selected the concentration of AZD5363 and rituximab at 40 μmol/L and 50 μg/mL for further study, and divided into 4 groups:control group, AZD5363 group, rituximab group and combined treatment group (AZD5363 plus rituximab),and the expression of p-Akt and T-Akt was detected by western blot(WB) after 24 hours.4. Use SPSS statistical 19.0 software to analyze data.Results 1. With the increasing of the rituximab concentrations (25.50. 75.100 ug/mL), growth inhibition rate was increased slightly in Daudi cells after 24 hours (16.35%、18.15%、21.89、24.36%).2. With the increasing of the AZD5363 concentrations (5.10.20.40.80μmol/L), growth inhibition rate were increased in Daudi cells after 24 hours (5.61%、13.42%、18.02%、21.32%, 55.23%).3. Growth inhibition rate of the combination group were 35.13%、 44.73%、71.33% respectively after 24.48.72 hours, which was significantly improved cytotoxicity when compared with AZD5363 or rituximab (P<0.05). 4. Apoptosis rate in combination groupsafter 48 hours (47.21%) was statistically higer than AZD5363 group(29.17%) and rituximab group(19.96%)(P<0.05); apoptosis rate in combination groups after 72 hours (65.23%)was also statistically higer than AZD5363 group(40.70%) and rituximab group(22.61%)(P<0.05).5. The expression of p-Akt in combination group was greatly lower (0.18±0.04) than alone AZD5363 (0.33±0.03) or rituximab group (0.45±0.04) (P<0.05), while the change of the expression of T-Akt was not obvious among the combination groups, the AZD5363 group and the rituximab group (P>0.05).Conclusion AZD5363 has a moderate effect on growth inhibition and apoptosis of Daudi cells, while the effects of rituximab are weaker. The anti-tumor effect of Daudi cells is significantly enhanced when AZD5363 in combination with rituximab, and its action mechanism is associated with the deserasing expression of P-Akt protein.