线粒体铁蛋白对顺铂抑制人肺癌细胞A549/DDP增殖的作用机制

Effect of ferritin mitochondrial on the proliferation of human lung cancer cell A549/DDP

目的探讨线粒体铁蛋白(FTMT)通过hepcidin-BMP6-SMAD7信号通路对顺铂(DDP)抑制人肺癌细胞A549/DDP增殖的作用。方法实验分为0(正常RPMI-1640培养基),1.0,2.5,5.0,7.5和10μg·mL^(-1)DDP组(1.0,2.5,5.0,7.5和10μg·mL^(-1)DDP培养基处理)。以噻唑蓝(MTT)法检测A549和A549/DDP在DDP作用后的细胞存活率;以流式细胞术检测细胞凋亡、活性氧(ROS)和线粒体膜电位(MMP)的水平;以逆转录聚合酶链反应(qPCR)法检测细胞中FTMT、铁蛋白重链(FTH)、铁蛋白轻链(FTL)基因的表达水平。结果DDP诱导后细胞的存活率随着药物浓度的增加而显著降低。0和10μg·mL^(-1)DDP组A549/DDP细胞早期凋亡率分别为(1.75±0.43)%和(0.95±0.28)%,坏死率分别为(2.35±0.42)%和(7.40±0.49)%,ROS水平分别为(4.25±0.49)%和(12.31±0.96)%,MMP水平分别为(89.00±2.38)%和(82.40±0.95)%,差异均有统计学意义(均P<0.05)。随着DDP浓度的增加,A549/DDP细胞中的Bax、caspase3、FTMT、铁调素、转铁蛋白受体(TfR1和TfR2)、BMP6、SMAD7和hjv的mRNA水平先升高后降低,Bcl-2和储铁蛋白(FTH和FTL)的mRNA水平先降低后升高。结论FTMT的高表达与DDP的耐药有一定关系,FTMT-hepcidin-BMP6-SMAD7信号通路参与了DDP诱导人肺癌A549/DDP细胞的凋亡过程。

Objective To investigate the effect of ferritin mitochondrial(FTMT)on the proliferation of human lung cancer cell A549/DDP via hepcidin-BMP6-SMAD7 signaling pathway.Methods The experiment was divided into 0(normal RPMI-1640 medium),1.0,2.5,5.0,7.5 and 10μg·mL^(-1)cisplatin(treated with 1.0,2.5,5.0,7.5 and 10μg·mL^(-1)cisplatin)groups.The survival rate of A549 and A549/DDP cells were measured by MTT assay after cisplatin induced.Flow cytometry was employed to assess the apoptosis,reactive oxygen species(ROS),and mitochondrial membrane potential(MMP)levels of the treated cells.The expression of several genes were measured by the qPCR,such as FTMT,ferritin heavy chain(FTH),ferritin light chain(FTL).Results The survival rates of A549/DDP cells were obviously decreased after cisplatin induced.The early apoptosis rate of A549/DDP cells in 0 and 10μg·mL^(-1)cisplatin groups were(1.75±0.43)%and(0.95±0.28)%,the necrosis rate were(2.35±0.42)%and(7.40±0.49)%,the ROS levels were(4.25±0.49)%and(12.31±0.96)%,the MMP levels were(89.00±2.38)%and(82.40±0.95)%,all with significant difference(all P<0.05).With the increase of cisplatin dosage,the mRNA levels of Bax,caspase3,FTMT,hepcidin,transferrin receptor(TfR1 and TfR2),bone morphogenetic protein(BMP6),SMAD7 and hemojuvelin(Hjv)in A549/DDP cells increased first and followed by a decrease,while the mRNA levels of Bcl-2,ferritin(FTH and FTL)decreased first and followed by an increase.Conclusion The high expression of FTMT was associated with DDP resistance,and the FTMT-hepcidin-BMP6-SMAD7 signaling pathway was involved in cisplatin induced apotosis of human lung cancer cells A549/DDP.