土荆皮乙酸诱导鼻咽癌细胞铁死亡的作用及机制

Role and Mechanism of Ferroptosis in Nasopharyngeal Carcinoma Cells Induced by Pseudolaric Acid B

目的探讨土荆皮乙酸诱导鼻咽癌发生铁死亡的机制。方法CCK-8法检测土荆皮乙酸(pseudolaric acid B,PAB)干预鼻咽癌细胞的细胞毒性。将鼻咽癌细胞分为对照组、土荆皮乙酸组(PAB组)、土荆皮乙酸+铁死亡抑制剂组(PAB+Fer-1组),划痕实验检测土荆皮乙酸对鼻咽癌细胞迁移能力的影响;化学发光法检测鼻咽癌细胞丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽(GSH)含量水平;Western Blot检测鼻咽癌细胞铁死亡相关蛋白谷胱甘肽过氧化物酶4(GPX4)、重肽铁蛋白(FTH1)、溶质载体家族7成员11(SLC7A11)、环氧化酶2(COX-2)的表达水平;流式细胞术检测鼻咽癌细胞的活性氧(ROS)水平。结果土荆皮乙酸随着剂量的增加和时间的延长抑制鼻咽癌细胞的细胞活力并诱导细胞死亡的作用增强。土荆皮乙酸能够明显抑制细胞迁移和增殖,促进细胞凋亡。土荆皮乙酸增加ROS水平、降低GSH水平、降低SOD活性、增加MDA浓度;下调GPX4、FTH1、SLC7A11表达,上调COX-2表达。土荆皮乙酸+铁死亡抑制剂组降低ROS水平、MDA浓度,增加GSH、SOD水平,上调GPX4、FTH1、SLC7A11蛋白表达,下调COX-2的表达。结论天然产物土荆皮乙酸通过促进鼻咽癌细胞凋亡及抑制细胞增殖和迁移来发挥抗肿瘤作用,其机制可能与诱导细胞铁死亡有关。

Objective This study investigated the mechanism study of ferroptosis induced by pseudolaric acid B(PAB)in nasopharyngeal carcinogenesis.Methods CCK-8 assay was used to detect the cytotoxicity of PAB intervention in nasopharyngeal carcinoma cells.The nasopharyngeal carcinoma cells were divided into control group,PAB group and PAB+ferroptosis inhibitor group(PAB+Fer-1 group),and the effect of PAB on the migration ability of nasopharyngeal carcinoma cells was detected by scratch assay.The levels of malondialdehyde(MDA),superoxide dismutase(SOD)and glutathione(GSH)in nasopharyngeal carcinoma cells were detected by chemiluminescence.The ferroptosis-related proteins in nasopharyngeal carcinoma cells were detected by Western Blot.The expression levels of glutathione peroxidase 4(GPX4),FTH1,SLC7A11 and cyclooxygenase-2(COX-2)in nasopharyngeal carcinoma cells were detected by Western Blot.The reactive oxygen species(ROS)levels in nasopharyngeal carcinoma cells were detected by flow cytometry.Results PAB inhibited cell viability and induced cell death in nasopharyngeal carcinoma cells with increasing dose and time.It significantly inhibited cell migration and proliferation and promoted apoptosis.PAB increased ROS level,decreased the GSH level and SOD activity and increased MDA concentration,down-regulated the expressions of GPX4,FTH1 and SLC7A11 and up-regulated COX-2 expression.The decrease of ROS level,MDA concentration,the increase of GSH level and SOD activity,up-regulation of protein expressions of GPX4,FTH1 and SLC7A11 and down-regulation of COX-2 expression in the group of PAB+ferroptosis inhibitor.Conclusion The natural product PAB exerts anti-tumor effects by promoting apoptosis and inhibiting cell proliferation and migration in nasopharyngeal carcinoma cells,and its mechanism may be related to the induction of cellular ferroptosis.