Akt抑制剂与利妥昔单抗联合应用对伯基特淋巴瘤细胞株Daudi增殖及凋亡的影响

摘要

Objective To investigate the effect of Akt inhibitor (AZD5363) combined with rituximab on cell prolifer-ation and apoptosis of Burkitt′s lymphoma (BL) Daudi cells in vitro, and to explore its mechanism.Methods Human BL Daudi cells were cultured and then were divided into four groups:the control group (treated by 1640 culture medium with fetal bovine serum) , AZD5363 group (treated by 40 μmol/L AZD5363) , rituximab group (treated by 50 μg/mL ritux-imab) and the combination group (treated by 40 μmol/L AZD5363 +50 μg/mL rituximab).They were all put into the incubator for 24-72 h.The growth inhibition and apoptosis rate of Daudi cells were detected by CCK-8 colorimetry and flow cytometer respectively.In addition, the expression of phosphorylated Akt (p-Akt) and total Akt (T-Akt) was detected by Western blotting.Results The growth inhibition rate of the combination group after 24, 48 and 72 h was significantly in-creased as compared with that of the AZD5363 and rituximab groups (all P＜0.05).The difference of apoptosis rate among the control group and AZD5363 group, rituximab group and the combination group was statistically significant after 48 and 72 h, and the apoptosis rate in the combination group was higher than those of the AZD 5363 and rituximab group after 48 and 72 h (all P＜0.05).The expression of p-Akt in the control group, AZD5363 group, rituximab group and the combina-tion group were respectively 0.59 ±0.06, 0.33 ±0.03, 0.45 ±0.04 and 0.18 ±0.04 after 24 h, and the difference of ex-pression of p-Akt was statistically significant among the control group and experimental groups , between AZD5363 group and the combination group, between the combination group and AZD5363 group, rituximab group (all P＜0.05).Conclu-sions AZD5363 in combination with rituximab can significantly inhibit cell proliferation and promote the apoptosis of BL Daudi cells.Inhibition of Akt phosphorylation and blocking of PI 3k/Akt signal transduction pathway may be the mechanism of its antitumor effect .%目的 探讨Akt抑制剂(AZD5363)与利妥昔单抗联合应用对伯基特淋巴瘤(BL)细胞株Daudi增殖和凋亡的影响,并探讨其相关机制.方法 培养人BL细胞株Daudi,将其分为对照组(只含等量溶媒)、AZD5363组(加入40μmol/L AZD5363)、利妥昔单抗组(加入50μg/mL利妥昔单抗)、联合用药组(加入40μmol/L AZD5363+50μg/mL利妥昔单抗),置于培养箱中培养24～72 h.分别采用CCK8法、流式细胞术测算AZD5363、利妥昔单抗及两者联合对Daudi细胞的增殖抑制和凋亡率,采用Western blotting法检测细胞磷酸化Akt(p-Akt)和总Akt(T-Akt)蛋白.结果 与AZD5363组、利妥昔单抗组相比,联合用药组在24、48、72 h时Daudi细胞增殖抑制率高(P均＜0.05).AZD5363组、利妥昔单抗组、联合用药组与对照组相比,联合用药组与AZD5363组、利妥昔单抗组相比,干预Daudi细胞48、72 h后细胞凋亡率高(P均＜0.05).对照组、AZD5363组、利妥昔单抗组、联合用药组细胞p-Akt相对表达量分别为0.59±0.06、0.33±0.03、0.45±0.04、0.18±0.04,AZD5363组、利妥昔单抗组、联合用药组与对照组相比,联合用药组与AZD5363组、利妥昔单抗组相比,P均＜0.05.结论 AZD5363与利妥昔单抗联合应用可增强对BL细胞株Daudi的增殖抑制作用,促使细胞凋亡,其作用机制可能是通过抑制Akt磷酸化,阻断PI3k/Akt信号转导通路,从而发挥抗肿瘤效应.