目的 探讨淫羊藿苷(icariin，ICA)对人口腔鳞癌细胞系CAL27细胞铁死亡的影响及其可能的作用机制。方法 用不同浓度ICA处理人口腔鳞癌细胞系CAL27，用CCK-8法检测细胞增殖情况以选取最佳浓度;用不同浓度铁死亡抑制剂处理CAL27细胞，用CCK-8法检测铁死亡抑制剂对细胞的毒性作用以选取最佳浓度;将CAL27细胞随机分为对照组、ICA组和ICA联合铁死亡抑制剂组。用Transwell小室实验检测细胞迁移和侵袭能力;检测各组细胞超氧化物歧化酶(superoxide dismutase，SOD)、丙二醛(malondialdehyde，MDA)和谷胱甘肽(glutathione，GSH)水平;用实时荧光定量聚合酶链式反应(quantitative real time polymerase chain reaction，RT-PCR)检测细胞铁死亡影响因子重链亚基溶质载体家族 7成员 11(solute vector family 7 member 11，SLC7A11)、谷胱甘肽过氧化物酶4(glutathione peroxidase 4，GPX4)和铁蛋白重链(ferritin heavy chain，FTH1)mRNA的表达情况;用蛋白印迹法(Western blotting)检测c-Jun氨基末端激酶(c-Jun amino terminal kinase，JNK)和p53蛋白(protein 53，p53)的表达情况。结果 CAL27细胞增殖能力随着ICA和铁死亡抑制剂浓度的升高而降低(P＜0。05);与对照组比较，ICA组CAL27细胞侵袭数量、细胞迁移数量、SOD、GSH、SLC7A11和GPX4 mRNA的表达水平降低，MDA和FTH1 mRNA、JNK和p53蛋白的表达水平升高(P＜0。05);与ICA组比较，ICA联合铁死亡抑制剂组CAL27 细胞侵袭数量、细胞迁移数量、SOD、GSH、SLC7A11和GPX4 mRNA的表达水平升高，MDA、FTH1 mRNA表达水平、JNK和p53蛋白的表达水平降低(P＜0。05)。结论 ICA对口腔鳞癌细胞CAL27增殖、侵袭和迁移能力有一定的抑制作用，可能是通过激活JNK/p53通路提高细胞内铁死亡相关因子水平，降低细胞抗氧化能力发挥作用的。
Effect of icariin on iron death in human oral squamous cell carcinoma cell line CAL27
Objective To investigate the effect of icariin(ICA)on iron death in human oral squamous cell carcinoma(OSCC)cell line CAL27 and to explore its possible mechanism.Methods Treated with different concentration ICA,the CAL27 cell proliferation was detected by CCK-8 method.Treated with different concentration iron death inhibitors,the toxic effect of iron death inhibitors on cells was detected by CCK-8 method.The optimal concentration was selected.The OSCC cell line CAL27 were randomly divided into control group,ICA group,and ICA combined with iron death inhibitors group.The ability of cell migration and invasion was detected by using Transwell chamber test.The activity of superoxide dismutase(SOD),the content of malondialdehyde(MDA)and the level of glutathione(GSH)were measured.The expression of heavy chain subunit solute vector family 7 member 11(SLC7A11),glutathione peroxidase 4(GPX4),ferritin heavy chain(FTH1)mRNA were detected by using RT-PCR.The proteins expression level of c-Jun amino terminal kinase(JNK)and p53 were detected by Western blotting.Results The proliferation ability of CAL27 cells decreased with the increase of the concentration of ICA and iron death inhibitor(P＜0.05).Compared with the control group,the invasion number,cell migration number and the levels of SOD,GSH,SLC7A11 and GPX4 mRNA expression were decreased,and the levels of MDA,FTH1 mRNA expression,JNK and p53 protein expression were increased in ICA group(P＜0.05).Compared with ICA group,the invasion number,cell migration number and the levels of SOD,GSH,SLC7A11 and GPX4 mRNA expression were increased,and the levels of MDA,FTH1 mRNA expression,JNK and p53 protein expression were decreased in ICA combined with iron death inhibitors group(P＜0.05).Conclusion ICA can inhibit the proliferation,invasion and migration of OSCC cell line CAL27,which may play a role by activating JNK/p53 pathway,increasing the level of intracellular iron death related factors and reducing the antioxidant capacity of cells.
万方数据
维普
