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目的:探讨蛋白激酶B(又称Akt)抑制剂AZD5363对人乳腺癌MDAMB-231细胞增殖、迁移和凋亡的影响,并探讨其可能的分子作用机制。方法:用不同浓度(0.5、1、5、10、20和50μmol/L)的AZD5363处理MDA-MB-231细胞后,采用MTT法检测细胞增殖活力,FCM法分析细胞周期的变化,划痕愈合实验和Transwell小室法检测细胞迁移能力的变化,TUNEL法测定细胞凋亡率,蛋白质印迹法检测细胞周期和凋亡相关蛋白的表达水平变化。结果:AZD5363可抑制MDA-MB-231细胞的增殖活力(P<0.05),并呈剂量依赖性;AZD5363可通过上调p53表达和下调cyclin B1表达(P值均<0.05),阻滞细胞周期于S期(P<0.05)。AZD5363能够明显抑制MDAMB-231细胞的迁移(P<0.05),同时诱导MDA-MB-231细胞发生凋亡(P<0.05),其分子机制可能与AZD5363促进caspase-3和多聚ADP-核糖聚合酶[poly(ADP-ribose)polymerase,PARP]的剪切活化相关(P值均<0.05)。结论:AZD5363能够抑制细胞增殖和迁移,同时诱导MDA-MB-231细胞凋亡,从而表现出抗肿瘤活性。
Objective: To investigate the effect of AZD5363, an inhibitor of protein kinase B (Akt), on the proliferation, migration and apoptosis of human breast cancer MDAMB-231 cells and to explore its possible molecular mechanism. Methods: MDA-MB-231 cells were treated with different concentrations of AZD5363 (0.5, 1, 5, 10, 20 and 50 μmol / L). Cell viability was measured by MTT assay. Cell cycle changes were analyzed by FCM. The changes of cell migration were detected by Transwell chamber assay, the apoptosis rate was measured by TUNEL method, and the expression of cell cycle and apoptosis related protein was detected by Western blotting. RESULTS: AZD5363 could inhibit the proliferation of MDA-MB-231 cells in a dose-dependent manner (P <0.05). AZD5363 could up-regulate the expression of p53 and down-regulate the expression of cyclin B1 (all P <0.05) S phase (P <0.05). AZD5363 could significantly inhibit the migration of MDAMB-231 cells (P <0.05) and induce apoptosis of MDA-MB-231 cells (P <0.05). The molecular mechanism of AZD5363 may be related to the promotion of caspase-3 and ADP- (ADP-ribose) polymerase, PARP] (all P <0.05). Conclusion: AZD5363 can inhibit cell proliferation and migration as well as induce apoptosis of MDA-MB-231 cells and thus show antitumor activity.