检验医学与临床2025，Vol.22Issue(18)：2498-2503,6.DOI:10.3969/j.issn.1672-9455.2025.18.009
ALOX15调控GPX4介导铁死亡抑制胶质瘤细胞增殖、迁移和侵袭
ALOX15 regulates GPX4-mediated ferroptosis to suppress proliferation,migration and invasion in glioma cells
摘要
Abstract
Objective To investigate the effect of arachidonic acid 15-lipoxygenase(ALOX15)regulating glutathione peroxidase 4(GPX4)-mediated ferroptosis on the proliferation,migration and invasion of glioma cells.Methods Glioma cell(A172,U87,LN229)and normal human astrocytes(HA1800)were cultured.ALOX15 mRNA and protein expression were detected by quantitative real-time PCR(qRT-PCR)and West-ern blot.The cell line with the lowest ALOX15 mRNA and protein expression was divided into three groups:control group(normal culture without treatment),ALOX15 group(transfected with pcDNA-ALOX15),ALOX15+GPX4 group(co-transfected with pcDNA-ALOX15 and pcDNA-GPX4).Cell proliferation was measured by CCK-8 assay,migration by scratch assay,and invasion by Transwell chamber assay.Intracellular reactive oxygen species(ROS)level was detected using DCFH-DA fluorescent probe.Ferroptosis-related pro-teins-glutathione peroxidase 4(GPX4),long-chain acyl-CoA synthetase 4(ACSL4),NADPH oxidase 1(NOX1)and cyclooxygenase-2(COX2)were analyzed using qRT-PCR and Western blot.Results ALOX15 mRNA and protein expression were lower in glioma cells A172,U87,and LN229 than in normal human astro-cytes HA1800(P＜0.05);compared with glioma cells A172,ALOX15 mRNA and protein were lower in glio-ma cells U87(P＜0.05);compared with glioma cells U87,ALOX15 mRNA and protein were higher in glioma cells LN229(P＜0.05).U87 cells showed the lowest ALOX15 mRNA and protein levels,and were therefore used for subsequent experiments.Compared with the control group,ALOX15 mRNA and protein expression was higher in the ALOX15 group(P＜0.05),while cell proliferation,migration and invasion rates were de-creased(P＜0.05).Compared with the control group,ROS level,ACSL4,NOX1 and COX2 mRNA and pro-tein expression was higher in ALOX15 group(P＜0.05),while GPX4 mRNA and protein expression was lower(P＜0.05).Compared with the ALOX15 group,GPX4 mRNA and protein expression was higher in the ALOX15+GPX4 group(P＜0.05),proliferation,migration and invasion rates were increased(P＜0.05),while ROS level,ACSL4,NOX1 and COX2 mRNA and protein expression were lower(P＜0.05).Conclusion Overex-pression of ALOX15 suppresses the proliferation,migration,and invasion of glioma cells,possibly by down-regulating GPX4 and thereby inducing ferroptosis.关键词
花生四烯酸15-脂氧合酶/谷胱甘肽过氧化物酶4/长链酯酰辅酶A合成酶4/铁死亡/胶质瘤Key words
arachidonic acid 15-lipoxygenase/glutathione peroxidase 4/long-chain acyl-CoA syn-thetase 4/ferroptosis/glioma分类
医药卫生引用本文复制引用
邹和琴,何娟,范冶..ALOX15调控GPX4介导铁死亡抑制胶质瘤细胞增殖、迁移和侵袭[J].检验医学与临床,2025,22(18):2498-2503,6.基金项目
湖南中医药大学校院联合基金项目(2024XYLH224). （2024XYLH224）
