摘要
目的:探讨磷脂酰肌醇3磷酸激酶(PI3K)雷帕霉素靶蛋白(m TOR)激酶(PI3K/m TOR)抑制剂GDC-0980对人肺腺癌细胞株A549细胞的诱导凋亡作用以及作用机制。方法 :选择不同浓度GDC-0980作用于A549细胞48h后,通过CCK8法检测GDC-0980对A549细胞增殖的影响,应用酶联免疫吸附法(ELISA)检测细胞凋亡内标志物核小体的形成,采用Annexin V-FITC/PI双染法流式细胞术以及免疫印迹法检测细胞凋亡。结果:GDC-0980能有效抑制A549细胞的增殖(66.61±5.72%),浓度越高抑制作用越强,抑制率可达66.61%。Annexin V-FITC/PI双染法染色显示,A549细胞膜发生翻转、破裂,最终导致凋亡。GDC-0980各浓度组凋亡率均显著增加,且随着GDC-0980浓度的增加,凋亡率也逐渐增加。结论:PI3K/m TOR抑制剂GDC-0980具有机制肺腺癌细胞株A549细胞生长、促凋亡的作用,并呈浓度依赖关系。
Objective:To study the effects of phosphatidylinositide 3-kinases/mammalian target of rapamycin kinase inhibitor GDC-0980 on induction of apoptosis and its possible mechanisms in human glioma A549 cells. Methods:The A549 cells were treated with different concentrations of GDC-0980 for 48 hrs. The effects of BKM on cell proliferation were detected by CCK8 kit. The nucleosome was detected by ELISA kit. The cell apoptosis was detected by Annexin V-FITC/PI staining. Results:The resulting form CCK8 test suggested that GDC-0980 can inhibit A549 proliferation in a con-centration dependent manner and the maximum inhibitory rate is 66.61%(66.61 ±5.72%). ELISA showed the change of A549 cell morphology including nucleosome after incubation with GDC-0980. The result of flow cytometry detected showed that A549 cells apoptosis ratio gradually increased when the concentration of GDC-0980 increased. Conclusion:GDC-0980 can inhibit the proliferation of A549 cells significantly. GDC-0980 can induce apoptosis of A549 cells in a concen-tration dependent manner.
出处
《交通医学》
2015年第4期326-328,332,共4页
Medical Journal of Communications