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目的:研究1,8-二甲基-3,5,7-三硝基-2-喹诺酮对人肺癌A549细胞的促凋亡作用及其可能的分子机制。方法:四甲基偶氮唑蓝(MTT)比色法测定1,8-二甲基-3,5,7-三硝基-2-喹诺酮对A549细胞增殖的影响;流式细胞术检测细胞凋亡率;蛋白免疫印迹法(Western blot)检测1,8-二甲基-3,5,7-三硝基-2-喹诺酮对A549细胞中活化型半胱天冬酶-3(cleavedCaspase-3),B细胞淋巴瘤-2(Bcl-2),Bcl-2相关X蛋白(Bax),总蛋白激酶B(Akt),磷酸化蛋白激酶B(p-Akt),叉头框蛋白O1(FOXO1),磷酸化叉头框蛋白O1(p-FOXO1),Bcl-2促细胞凋亡蛋白(Bim)表达的干预作用。结果:1,8-二甲基-3,5,7-三硝基-2-喹诺酮(1.25,2.5,5,10,20 mg·L-1)能质量浓度和时间依赖性地抑制A549细胞增殖。作用12,24 h的半数抑制浓度(IC50)分别为2.80,2.05μmol·L-1。流式细胞术结果显示1,8-二甲基-3,5,7-三硝基-2-喹诺酮能显著诱导细胞凋亡,随着药物质量浓度的提高,细胞凋亡率逐渐上升。Western blot检测显示,1,8-二甲基-3,5,7-三硝基-2-喹诺酮上调cleaved-Caspase-3和Bax蛋白的表达,下调Bcl-2蛋白的表达。同时会下调p-Akt和p-FOXO1的表达,上调Bim的表达。结论:1,8-二甲基-3,5,7-三硝基-2-喹诺酮能够诱导A549细胞凋亡,该药理作用可能与1,8-二甲基-3,5,7-三硝基-2-喹诺酮抑制磷脂酰肌醇3-激酶(PI3K)/Akt/FOXO1信号通路有关。
AIM: To investigate the apoptosis-promoting effect of 1,8-dimethyl-3,5,7-trinitro-2-quinolone on human lung cancer cell line A549 and its possible molecular mechanism. Methods: MTT assay was used to determine the effect of 1,8-dimethyl-3,5,7-trinitro-2-quinolone on the proliferation of A549 cells. Flow cytometry Apoptosis rate; Western blot was used to detect the effect of 1,8-dimethyl-3,5,7-trinitro-2-quinolone on cleaved Caspase- 3, Bcl-2, Bcl-2, Akt, p-Akt, FOXO1, p-FOXO1 and Bcl-2. Results: 1,8-Dimethyl-3,5,7-trinitro-2-quinolone (1.25,2.5,5,10,20 mg · L-1) inhibited A549 cells in a concentration-and time- proliferation. The half-maximal inhibitory concentration (IC50) at 12 and 24 h were 2.80 and 2. 05 μmol·L -1, respectively. The results of flow cytometry showed that 1,8-dimethyl-3,5,7-trinitro-2-quinolone could significantly induce apoptosis. With the increase of the drug concentration, the apoptosis rate gradually increased. Western blot showed that 1,8-dimethyl-3,5,7-trinitro-2-quinolone upregulated cleaved-Caspase-3 and Bax protein expression and down-regulated Bcl-2 protein expression. At the same time, it down-regulated the expression of p-Akt and p-FOXO1 and up-regulated the expression of Bim. CONCLUSION: 1,8-Dimethyl-3,5,7-trinitro-2-quinolone can induce A549 cell apoptosis, which may be related to 1,8-dimethyl-3,5,7- Nitro-2-quinolones inhibit phosphoinositide 3-kinase (PI3K) / Akt / FOXO1 signaling.