论文部分内容阅读
目的分析磷脂酰肌醇3激酶(phosphatidylinositol 3 kinase,PI3K)/蛋白激酶B(protein kinase B,AKT)通路是否通过长细胞caspase-8样抑制蛋白(cellular FLICE inhibitory protein,c-FLIP-L)的介导来影响乳腺癌细胞对肿瘤坏死因子相关凋亡诱导配体(TNF related apoptosis inducing ligand,TRAIL)的敏感性,并探讨其机制。方法用40 nmol/L wortmannin(AKT活化的特异性抑制剂)、100 ng/ml TRAIL、40 nmol/L wortmannin+100 ng/ml TRAIL处理人乳腺癌MDA-MB-231细胞,并设空白对照组。MTT法检测药物作用24、48、72 h后的细胞增殖抑制率;流式细胞术检测药物作用48 h后的细胞凋亡率;Western blot法检测细胞中磷酸化AKT(pAKT)和c-FLIP-L的表达。结果TRAIL+wortmannin组24、48和72 h细胞增殖抑制率与其他组比较,均明显升高(P<0.05);TRAIL+wortmannin组与TRAIL组和wortmannin组比较,可明显促进细胞凋亡(P<0.05);随着wortmannin作用时间的延长,pAKT的表达水平明显降低,同时c-FLIP-L的表达水平也随之下降,而TRAIL对pAKT和c-FLIP-L的表达无明显影响。结论抑制PI3K/AKT通路可增强乳腺癌细胞对TRAIL的敏感性,这种作用可能是通过c-FLIP-L的介导来实现的。
OBJECTIVE: To determine if the phosphatidylinositol 3 kinase (PI3K) / protein kinase B (AKT) pathway is mediated by the cellular caspase-8 inhibitory protein (c-FLIP-L) Mediated to affect the sensitivity of breast cancer cells to tumor necrosis factor-related apoptosis inducing ligand (TRAIL), and to explore its mechanism. Methods Human breast cancer MDA-MB-231 cells were treated with 40 nmol / L wortmannin (100 μg / ml TRAIL, 40 nmol / L wortmannin + 100 ng / ml TRAIL) . MTT assay was used to detect the cell proliferation inhibition rate after 24, 48, and 72 h; the apoptosis rate was detected by flow cytometry after 48 h; the phosphorylation of AKT (pAKT) and c-FLIP -L expression. Results The inhibitory rates of cell proliferation in TRAIL + wortmannin group at 24, 48 and 72 h were significantly higher than those in other groups (P <0.05). Compared with TRAIL group and wortmannin group, the apoptosis of TRAIL + wortmannin group was significantly increased <0.05). With the prolongation of wortmannin time, the expression of pAKT was significantly decreased, while the expression of c-FLIP-L was also decreased. However, TRAIL had no significant effect on the expression of pAKT and c-FLIP-L. Conclusion Inhibition of PI3K / AKT pathway can enhance the sensitivity of breast cancer cells to TRAIL, and this effect may be mediated by c-FLIP-L.