CHAC1对肺癌细胞铁死亡的调控及其机制研究

Regulation of CHAC1 on iron death in lung cancer cells and its mechanism

目的本研究旨在探讨CHAC1对肺癌细胞铁死亡的调控及其作用机制。方法以人肺癌A549细胞为研究对象,通过外源性转染CHAC1的过量表达载体,调控CHAC1的表达,同时给予铁死亡诱导剂Erastin,通过四甲基偶氮唑盐比色法(MMT)法检测细胞铁死亡情况;通过细胞平板集落形成实验检测细胞铁死亡情况;通过流式细胞仪分析细胞周期。结果上调CHAC1的表达,可使肺癌细胞生长能力下降,与对照组细胞相比,具有统计学意义(P<0.05)。与转染空载体的对照细胞系相比,转染小干扰RNA细胞其克隆形成数明显下降(P<0.05)。与对照A549/对照细胞[G_0/G_1期(51.2±1.1)%;S期(36.4±0.8)%;G_2/M期(11.4±1.4)%]相比,稳定转染A549/CHAC1过表达和Erastin处理过的A549细胞G_1期的DNA含量明显增加,S期DNA含量明显减少(均P<0.05)。结论上调CHAC1的表达可显著增加肺癌细胞的铁死亡,抑制其增殖,其分子机制与上调CHAC1可使肺癌细胞停滞在G1期,阻止细胞进入S期有关。

Objective To investigate the regulation of CHAC1 on iron death in lung cancer cells and its mechanism of action.Methods Human lung cancer A549 cells were used as the research objects,and the expression of CHAC1 was regulated by exogenous transfection of CHAC1 overexpression vector.At the same time,iron death inducer Eratin was given to detect the iron death of cells by MMT method.Cell plate colony formation for monitoring cell lron death was detected experimentally;cell cycle was analyzed by flow cytometry.Results Up-regulating the expression of CHAC1 can reduce the growth ability of lung cancer cells,which was statistically significant compared with the control group cells(P<0.05).Compared with the control cell line transfected with the empty vector,the number of clone formation of transfected small interfering RNA cells was significantly reduced(P<0.05).Compared with control A549/control cells[G0/G1 phase(51.2±1.1)%;S phase(36.4±0.8)%;G2/M phase(11.4±1.4)%],A549/CHAC1 overexpressed and Eratin-treated A549 cells G1 phase DNA content increased significantly,and S phase DNA content decreased significantly(both P<0.05).Conclusion Up-regulating CHAC1 expression can significantly increase iron death of lung cancer cells and inhibit its proliferation.The molecular mechanism is related to up-regulating CHAC1 to arrest lung cancer cells in G1 phase and prevent cells from entering S phase.