摘要
目的 研究毛蕊异黄酮(calycosin, CA)调控糖原合成酶激酶-3β(glycogen synthase kinase-3β,GSK-3β)/核因子-E2相关因子2(nuclear factor erythroid-derived 2-like 2,Nrf2)/谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)铁死亡通路逆转结直肠癌5-氟尿嘧啶(5-fluorouracil, 5-FU)耐药。方法 选择5-FU耐药的结直肠癌细胞株HT-29/5-FU为实验对象,采用CCK8法检测5-FU、CA、5-FU+CA对细胞增殖的影响,计算逆转耐药倍数。采用克隆形成实验检测克隆形成数目,采用试剂盒检测结直肠癌细胞中ROS和MDA水平,采用Western blotting方法检测细胞中GSK-3β、Nrf2、GPX4的表达水平。结果 100μmol/L CA逆转HT-29/5-FU细胞对5-FU耐药,逆转倍数为2.65倍;CA、5-FU单用或联合作用于HT-29/5-FU细胞均能显著减少克隆形成以及Nrf2、GPX4表达,增加ROS、MDA水平及GSK-3β表达;与CA、5-FU单用比较,CA和5-FU联用进一步减少克隆形成以及Nrf2、GPX4表达,增加ROS、MDA水平及GSK-3β表达。结论 CA逆转结直肠癌细胞株5-FU耐药,其涉及的机制可能与调控GSK-3β/Nrf2/GPX4通路、激活铁死亡相关。
Objective To study the reversal effect of calycosin(CA)on 5-fluorouracil(5-FU)resistance of colorectal cancer by regulating the glycogen synthase kinase-3β(GSK-3β)/nuclear factor erythroid-derived 2-like 2(Nrf2)/glutathione peroxidase 4(GPX4)ferroptosis pathway.Methods Using 5-FU resistant colorectal cancer cell line HT-29/5-FU as the experimental subjects,CCK8 method was used to detect the effects of 5-FU,CA and 5-FU+CA on cell proliferation,and the reverse resistance multiple was calculated.The clones formation number was detected by colony formation assay,ROS and MDA levels in cells were detected by kit,and the expression levels of GSK-3β,Nrf2 and GPX4 in cells were detected by Western blotting.Results The resistance of HT-29/5-FU cells to 5-FU was reversed by 100μmol/L CA,the reversal ratio was 2.65 times.CA and 5-FU alone or in combination significantly decreased colony formation and the expression of Nrf2 and GPX4,and increased the levels of ROS,MDA and the expression of GSK-3βin HT-29/5-FU cells.Compared with CA and 5-FU alone,the combination of CA and 5-FU further reduced colony formation and the expression of Nrf2 and GPX4,and increased the levels of ROS,MDA and the expression of GSK-3β.Conclusion CA reverses 5-FU resistance in colorectal cancer cell lines,and its mechanism is related to regulation of GSK-3β/Nrf2/GPX4 pathway and activation of ferroptosis.
作者
梁家
刘俊杰
庞天舒
薛佳龙
刘德纯
LIANG Jia;LIU Junjie;PANG Tianshu;XUE Jialong;LIU Dechun(Department of Gastrointestinal Surgery,the First Affiliated Hospital of Henan University of Science and Technology,Luoyang 471000,China)
出处
《胃肠病学和肝病学杂志》
2025年第12期1792-1796,共5页
Chinese Journal of Gastroenterology and Hepatology
