芬太尼通过PI3K/AKT信号通路影响肺癌H1299细胞增殖与凋亡

Effects of fentanyl on proliferation and apoptosis of H1299 cells by modulating AKT activation

目的探讨芬太尼对人肺癌H1299细胞增殖、凋亡的影响。方法使用芬太尼(0.001、0.010、0.100、1.000μM)分别以12、24、48、72h对肺癌H1299细胞进行处理,通过CCK8法检测H1299细胞活力;DAPI核染色法检测细胞凋亡引起的核碎裂情况。Westernblotting检测凋亡相关蛋白Bcl-2和Bax的表达水平,Caspase-3活性检测试剂盒检测法检测H1299细胞Caspase-3的活性;Westernblotting检测p-AKT/AKT水平。结果芬太尼可以呈浓度、时间依赖抑制H1299细胞增殖能力。芬太尼(0.010、0.100、1.000μM)作用于人肺癌H1299细胞48h可显著促进其凋亡,差异有统计学意义(P<0.05)。同时,芬太尼(0.010、0.100、1.000μM)作用于人肺癌H1299细胞48h,可显著下调Bcl-2、上调Bax的表达水平,增加Caspase-3的活性,并且细胞的p-AKT/AKT水平显著降低。结论芬太尼可抑制人肺癌H1299细胞的增殖,促进人肺癌细胞的凋亡,其机制可能与抑制AKT的活化有关。

Objective To explored the effects of fentanyl on cell proliferation of H1299 cells. Methods After treating H1299 cells with different concentrations of fentanyl (0.001, 0.010, 0.100, 1.000 μM) for 12, 24, 48, 72 h, cell viability was detected by CCK-8 assay;the rate of cell apoptosis was determined by DAPI staining;the expression levels of Bax, Bcl-2, p-AKT and AKT protein were measured by Western blotting;Caspase-3 activity was determined by Caspase-3 activity assay kit. Results Compared with the control group, fentanyl obviously inhibited the viability of H1299 cells in a dose and time dependent way. Moreover, treatment with different concentrations of fentanyl(0.001, 0.010, 0.100, 1.000 μM) for 12, 24, 48, 72 h, the apoptosis rate of H1299 cells were significantly increased. The level of Bcl-2 protein reduced the level of Bax protein, and the activity of Caspase-3 in H1299 cells were increased after treatment with fentanyl (0.010, 0.100, 1.000 μM) for 48 h. Furthermore, fentanyl markedly inhibited p-AKT/AKT activity of H1299 cells. Conclusions Fentanyl can inhibit cell proliferation and promote cell apoptosis of human lung cancer, and its mechanism may be related to inhibition of AKT activation.