FBXW7通过抑制c-Myc/SOX2/SLC7A11促进头颈鳞癌细胞铁死亡

FBXW7 promotes ferroptosis in head and neck squamous cell carcinoma cells through inhibiting c-Myc/SOX2/SLC7A11

目的探究FBXW7对头颈鳞状细胞癌铁死亡的影响。方法体外培养头颈鳞癌细胞系HN4和HN6,构建FBXW7,SOX2过表达质粒,将质粒稳定转染细胞系培养,采用qRT-PCR和Western blot实验分别在转录水平和蛋白水平验证过表达转染效率。通过检测丙二醛(MDA)、谷胱甘肽(GSH)和活性氧(ROS)水平来验证过表达FBXW7后头颈鳞癌细胞的脂质过氧化水平。用铁死亡抑制剂Fer-1处理细胞后进一步检测细胞活力变化验证FBXW7对铁死亡的影响。通过Western blot检测转染过表达质粒后对细胞通路的影响。结果HN4和HN6细胞系在过表达FBXW7后表现出脂质过氧化水平增加,铁死亡抑制剂Fer-1能够有效逆转过表达FBXW7引起的细胞铁死亡。Western blot实验结果显示过表达FBXW7降低了c-Myc、SOX2和SLC7A11的表达。结论FBXW7通过降解c-Myc来调控SOX2-SLC7A11的表达,从而有效调控头颈鳞状细胞癌铁死亡。

Objective To explore the effect of FBXW7 on ferroptosis in head and neck squamous cell carcinoma.Methods Head and neck squamous cell lines HN4 and HN6 were cultured in vitro.FBXW7 and SOX2 overexpression plasmids were constructed,and the plasmids were stably transfected into cell lines.The overexpression transfection efficiency was verified at the transcription level and protein level by qRT-PCR and Western blot experiments,respectively.The lipid peroxidation levels of head and neck squamous cell carcinoma cells with overexpressing FBXW7 were verified by measuring malondialdehyde(MDA),glutathione(GSH),and reactive oxygen species(ROS)levels.After treating cells with ferroptosis inhibitor Fer-1,the changes in cell viability were further detected to verify the effect of FBXW7 on ferroptosis.The effect of transfection of the overexpressed plasmid on cellular pathways was detected by Western blot.Results HN4 and HN6 cell lines showed increased levels of lipid peroxidation after overexpression of FBXW7,and the ferroptosis inhibitor Fer-1 was able to effectively reverse the ferroptosis induced by overexpression of FBXW7.Western blot assay results showed that overexpression of FBXW7 reduced the expression of c-Myc,SOX2 and SLC7A11.Conclusion FBXW7 regulates the expression of SOX2-SLC7A11 by degrading c-Myc,thereby effectively regulating ferroptosis in HNSCC.