AZD1480靶向阻断JAK2-STAT3信号通路抑制胶质瘤细胞增殖研究

Targeted blockade of JAK2-STAT3 signaling pathway with AZD1480 suppressing growth in human glioma cell

目的探讨AZD1480对胶质瘤细胞增殖的靶向抑制作用及其机制。方法用不同浓度的AZD1480在不同时间段内处理胶质瘤细胞U87MG,然后分别采用Western Blot检测JAK2-STAT3通路活化、CCK-8检测细胞增殖、AnnexinⅤ/PI染色检测细胞凋亡,统计学分析AZD1480的影响。结果用不同浓度的AZD1480处理24 h后,U87MG细胞中的磷酸化活化蛋白p-JAK2的表达随浓度增加而逐渐降低,p-STAT3的表达则被完全抑制。与用0μM AZD1480分别处理24、48、72 h结果(0.657±0.037、1.359±0.051、1.919±0.092)相比,用0.5μM的AZD1480处理后,U87MG的细胞活性分别为0.554±0.028、1.127±0.053和1.359±0.125,差异有统计学意义(P<0.05);随着AZD1480浓度增加及干预时间延长,U87MG细胞活性的降低更为明显(P<0.05)。与对照组(0μM)比较,用0.5μM AZD1480处理过的U87MG细胞凋亡比例从(4.54±0.50)%增长至(6.81±0.70)%,差异有统计学意义(P<0.05);随着AZD1480浓度增加,U87MG细胞凋亡比例呈逐渐上升趋势(P<0.05)。结论应用AZD1480能够靶向阻断JAK2-STAT3信号通路活化,有效抑制人脑胶质细胞增殖、诱导其凋亡,从而有望成为人脑胶质瘤基因靶向治疗的一种新靶点和新策略。

Objective To study the effect and its relevant mechanism of targeted suppression growth of human glioma cell by AZD1480.Methods Human glioma cell line U87 MG was treated with indicated concentrations of AZD1480 for 24 to 72 hours.The protein expression of Jak2,p-Jak2,STAT3,and p-STAT3 were detected by Western blot.The cellular viability was analyzed by CCK-8 assay.The apoptosis was evaluated by Annexin V/PI staining.Results The protein activation,Jak2 and STAT-3 of U87 MG were blocked by AZD1480,and the cellular viability was suppressed in a time-and dose-dependent manner in U87 MG cells,and the apoptosis was inducted(P<0.05).Conclusion AZD1480 treatment might effectively inhibit constitutive and stimulus-induced Jak2 and STAT3 phosphorylation in human glioma cell and lead to a decrease in cell proliferation and induction of apoptosis,indicating that pharmacological inhibition of the Jak2-STAT3 pathway by AZD1480 should be considered for study in the treatment of patients with glioma.