洛铂通过PERK/eIF2α/ATF4/CHOP信号通路调节肺癌细胞A549的作用机制

Loplatin regulates the mechanism of action of lung cancer cell A549 through PERK/eIF2α/ATF4/CHOP signaling pathway

目的探讨洛铂通过调节蛋白激酶R样内质网激酶(protein kinase R-like endoplasmic reticulum kinase,PERK)/真核生物起始因子2α(eukaryotic initiation factor 2α,eIF2α)/活化转录因子4(activating transcription factor 4,ATF4)/CCAAT/增强子结合蛋白同源蛋白(enhancer-binding protein homologous protein,CHOP)信号通路对肺癌细胞A549增殖、凋亡、迁移的影响。方法将A549细胞分为对照组、洛铂组(12μmol/L洛铂)、PERK抑制组(5μmol/L GSK2606414)及洛铂(12μmol/L)+PERK抑制组(5μmol/L GSK2606414)。通过MTT法、Transwell实验及流式细胞术检测细胞增殖、迁移、侵袭及凋亡;蛋白质印迹法检测细胞PERK/eIF2α/ATF4/CHOP信号通路相关蛋白水平。结果与对照组比较,洛铂组细胞24及48 h的吸光度值、细胞迁移数和侵袭数均下降,细胞凋亡率、磷酸化PERK/PERK、磷酸化eIF2α/eIF2α、ATF4以及CHOP蛋白水平均升高(P<0.01),PERK抑制组细胞24及48 h的吸光度值、细胞迁移数和侵袭数均升高,PERK/eIF2α/ATF4/CHOP信号通路相关蛋白水平降低(P<0.01)。与洛铂组比较,洛铂+PERK抑制组A549细胞增殖、迁移数以及侵袭数均升高,细胞凋亡率、PERK/eIF2α/ATF4/CHOP信号通路相关蛋白水平均下降(P<0.01)。结论洛铂可能通过激活PERK/eIF2α/ATF4/CHOP通路抑制A549细胞增殖、迁移以及侵袭,促进其凋亡。

Objective To investigate the effects of lobaplatin on proliferation,apoptosis and migration of lung cancer cell A549 by regulating protein kinase R-like endoplasmic reticulum kinase(PERK)/eukaryotic initiation factor 2α(eIF2α)/activating transcription factor 4(ATF4)/CCAAT/enhancer binding protein homologous protein(CHOP)signaling pathway.Methods A549 cells were divided into control group,lobaplatin group(12μmol/L lobaplatin),PERK inhibition group(5μmol/L GSK2606414)and lobaplatin(12μmol/L)+PERK inhibition group(5μmol/L GSK2606414).MTT method,Transwell experiment and flow cytometry were used to detect cell proliferation,migration,invasion,and apoptosis.Western blot was used to detect the protein levels of PERK/eIF2α/ATF4/CHOP signaling pathway.Results Compared with the control group,the absorbance value,cell migration number and invasion number in lobaplatin group were decreased at 24 h and 48 h,and the apoptosis rate,phosphrylated-PERK/PERK,phosphrylated-eIF2α/eIF2α,ATF4 and CHOP protein levels were increased(P<0.01).The absorbance value,cell migration number and invasion number in PERK inhibited group were increased at 24 h and 48 h,and the protein levels related to PERK/eIF2α/ATF4/CHOP signaling pathway were decreased(P<0.01).Compared with lobaplatin group,the proliferation,migration and invasion numbers of A549 cells in lobaplatin+PERK inhibitory group were increased,and the apoptosis rate and the protein levels of PERK/eIF2α/ATF4/CHOP signaling pathway were decreased(P<0.01).Conclusion Lobaplatin may inhibit the proliferation,migration and invasion of A549 cells and promote cell apoptosis by activating the PERK/eIF2α/ATF4/CHOP pathway.