硫利达嗪联合顺铂抑制肺癌A549及A549/DDP细胞的增殖

Thioridazine in combination with cisplatin inhibits the proliferation of lung cancer A549 and A549/DDP cells

目的:探讨硫利达嗪(thioridazine,THD)联合顺铂(cisplatin,DDP)对肺癌A549细胞及其DDP耐药A549/DDP细胞增殖和凋亡的影响,以及可能的作用机制。方法:THD、DDP及THD联合DDP处理A549和A549/DDP细胞后,应用CCK-8法检测各组细胞的增殖情况,FCM法检测细胞凋亡及细胞周期分布,JC-1染色法检测细胞线粒体膜电位变化,化学发光法检测细胞中caspase 8和caspase 9的活性,蛋白质印迹法检测细胞中cleaved caspase 3蛋白的表达水平。结果:THD联合DDP可显著抑制A549及A549/DDP细胞的增殖(P值均<0.01),2药的联合效应为相加作用。THD联合DDP作用A549细胞24、48和72 h的联合指数分别为1.01、1.01和1.03,作用A549/DDP细胞的联合指数分别为1.11、1.03和1.00。THD联合DDP作用后,A549和A549/DDP细胞的凋亡率均显著高于未进行药物处理的对照组(P值均<0.01),A549和A549/DDP细胞的细胞周期均被阻滞于G0/G1期(P值均<0.05),线粒体膜电位均低于对照组(P值均<0.01),caspase 8和caspase 9的活性及cleaved caspase 3蛋白的表达水平均高于对照组(P值均<0.05)。结论:THD联合DDP可抑制A549及A549/DDP细胞的增殖,其作用可能与促进细胞凋亡有关。

Objective： To explore the effects of thioridazine （THD） combined with cisplatin （DDP） on the proliferation and apoptosis of human cancer A549 cells and the chemoresistant A549/DDP cells and possible mechanism. lung theirMethods： After treatment with THD, DDP and the combination of THD and DDP, respectively, the proliferation of A549 and A549/DDP cells was determined by CCK-8 assay, the apoptosis and cell cycle distribution were determined by using FCM, the mitochondrial membrane potential was determined by JC-1 staining, the activities of caspase 8 and caspase 9 were detected by chemiluminiscence assay, and the expression level of cleaved caspase 3 protein was examined by Western blotting.Results： THD in combination with DDP inhibited the proliferation of A549 and A549/DDP cells （both P 〈 0.01）, and the combined effect was additive. The combination index （CI） values of THD in combination with DDP in treatment of A549 and A549/DDP cells for 24, 48 and 72 h were 1.01, 1.01 and 1.03, and 1.11, 1.03 and 1.00, respectively. The apoptotic rates of A549 and AS49/DDP cells treated with THD in combination with DDP were higher than those not treated with THD and DDP （as control group） （both P 〈 0.01）, the cell cycle of A549 and AS49/DDP cells was arrested at G0/G1 phase （both P 〈 0.05）, and the mitochondrial membrane potentials were lower than that of the control group （both P 〈 0.01）; while the activities of caspase 8 and caspase 9 and the expression level of cleaved caspase 3 in THD combined with DDP group were higher than those in the control group （all P 〈 0.05）. Conclusion： THD in combination with DDP can inhibit the proliferation of lung cancer A549 and A549/DDP cells, and this effect may be related to inducing apoptosis.