二氯乙酸钠联合顺铂诱导人肺腺癌A549细胞株凋亡的实验研究

目的研究二氯乙酸钠(Sodium dichloroacetate,DCA-Na)、顺铂(DDP)联合对人肺腺癌A549细胞株增殖和凋亡的影响及其作用机制。方法用MTT法检测DCA-Na、DDP应用对A549细胞增殖抑制作用的影响;流式细胞仪(Annexin V-FITC/PI法)检测DCA-Na、DDP单药及两药联合作用于A549细胞凋亡率的变化;用分光光度法检测DCA-Na作用于A549细胞后半胱氨酸天门冬氨酸蛋白酶(Caspase)-3、-8、-9蛋白的活性。结果DCA-Na、DDP单药组均对A549细胞增殖有抑制作用,且呈明显的剂量-时间依赖性。0.4、2μg/mL的DDP与37.5、75、150μg/mL的DCA-Na联合作用A549细胞24、48、72 h后的抑制率显著高于同浓度DDP单药组(P<0.05),且2μg/mL的DDP与75μg/mL的DCA联合在48 h表现为协同作用。流式细胞仪检测显示,A549细胞联合组凋亡率显著高于各单药组(P<0.05)。DCA-Na作用于A549细胞后,分别在12、24、48、72 h测得Caspase-3、-8、-9蛋白活性显著高于对照组(P<0.05)。结论 DCA-Na对人肺腺癌A549细胞的增殖有抑制作用,并且随着药物浓度增加、作用时间延长,其抑制作用也增加(呈剂量和时间依赖性)。一定浓度范围的DCA-Na和DDP联合作用于人肺腺癌A549细胞能够产生协同作用。DCA-Na可诱导人肺腺癌A549细胞凋亡,且与DDP联合后其诱导凋亡作用更为显著。

Study on apoptosis of human lung adenocarcinoma cell line-A549 induced by sodium dichloroacetate combined with cisplantin

Objective To explore the proliferation and apoptosis effects of dichloroacetate(DCA-Na) combined with DDP on human lung adenocarcinoma cell line-A549 in vitro and to investigate the possible underlying mechanisms.Methods The inhibitory effects of DCA-Na combined with DDP on proliferation of A549 cells were determined by MTT assay.A549 cells apoptosis was evaluated by FCM with Annexin V-FITC and PI.The activity of Caspase-3,Caspase-8,Caspase-9 of A549 cells,which were affected by DCA-Na at different time points,was evaluated by spectrophotometry.Results DCA-Na and DDP used alone could inhibit the proliferation of A549 cells,and the inhibition effect showed dose-time dependent relationship.The combination of DDP(0.4 μg/mL,2 μg/mL)with DCA-Na(37.5 μg/mL,75 μg/mL,150 μg/mL)significantly increased the inhibition effect of A549 cells,compared with DDP group(P<0.05).The combination of DCA-Na(75 μg/mL)and DDP(2 μg/mL)showed synergy at 48 h.The number of A549 cells which were treated by DCA-Na(75 μg/mL)and/or DDP(2 μg/mL)was obviously fewer than that of control group.FCM showed that the rate of apoptosis of A549 cells combination group was significantly higher than that in DCA-Na and(DDP group)(P<0.05).When A549 cells had been treated with DCA(37.5 μg/mL,75 μg/mL,150 μg/mL)for different time,the activity of Caspase-3,Caspase-8,Caspase-9 was significantly higher than those of control group(P<0.05).Conclusion DCA-Na could inhibit the proliferation of A549 cells in a dose-time dependent manner.DCA-Na and DDP in certain concentration has synergy effects.DCA-Na could induce the apoptosis of A549 cells,and the induction effect of DCA-Na combined with DDP is more significant.