热疗调控铁死亡通路对舌鳞癌细胞生物学行为影响的研究

Effect of hyperthermia on biological behavior of squamous cell carcinoma of the tongue by regulating ferroptosis pathway

目的探讨热疗(HT)对舌鳞癌细胞系CAL-27铁死亡的调控作用和可能机制。方法采用CCK-8法检测铁死亡抑制剂Fer-1的半抑制浓度(IC50)并用于后续实验。CAL-27细胞系按实验设计分为HT组、对照组、Fer-1组以及HT+Fer-1组。采用各自检测试剂盒检测活性氧水平、铁离子浓度,采用实时反转录PCR检测p53、转铁蛋白受体1(TfR1)的mRNA水平,细胞划痕法检测细胞迁移,流式细胞术检测细胞凋亡。结果热疗组CAL-27细胞系活性氧水平(P<0.01)和铁离子浓度(P<0.001)显著上调,p53及TfR1的mRNA表达量也明显增加(P值均<0.01),细胞迁移能力下降(P<0.001),凋亡率升高(P<0.01)。HT+Fer-1组与HT组相比活性氧水平(P<0.001)、铁离子浓度(P<0.001)、p53及TfR1的mRNA表达量均降低(P值均<0.01),细胞迁移能力恢复(P<0.01),凋亡率也降低(P<0.01)。结论热疗可能通过激活p53/TfR1通路诱导舌鳞癌细胞系CAL-27铁死亡,抑制其迁移能力并促进其凋亡。

Objective To investigate the regulation and possible mechanism of hyperthermia(HT)on the ferroptosis of squamous cell carcinoma of the tongue cell line CAL-27.Methods Half maximal inhibitory concentration(IC50)of Fer-1,an inhibitor of ferroptosis,was detected by CCK-8 assay and used for subsequent experiments.CAL-27 cells were divided into the HT,control,Fer-1 and HT+Fer-1 groups according to experimental design.Reactive oxygen species(ROS)levels and iron ion concentration were determined by corresponding detection kits.The p53 and TfR1 mRNA levels were detected by real-time reverse transcription PCR.Cell migration was detected by cell scratch test and cell apoptosis was detected by flow cytometry.Results HT significantly up-regulated the ROS levels(P<0.01)and iron ion concentration(P<0.001),and significantly increased the expression levels of p53 and TfR1 mRNA(both P<0.01).The cell migration ability was decreased(P<0.001),whereas cell apoptosis rate was increased by HT(P<0.01).In the HT+Fer-1 group,the ROS levels(P<0.001),iron ion concentration(P<0.001),expression levels of p53 and TfR1 mRNA(both P<0.01)were significantly down-regulated,the cell migration ability was recovered(P<0.01),and cell apoptosis rate was decreased(P<0.01)compared with those in the HT group,respectively.Conclusions HT may induce the ferroptosis of CAL-27 cell line,inhibit cell migration ability and promote cell apoptosis by activating the p53/TfR1 pathway.