青蒿琥酯通过介导铁死亡逆转结肠癌细胞5-氟尿嘧啶耐药的作用研究

Artesunate reverse the resistance of colon cancer cells to 5-fluorouracil by mediating ferroptosis

目的探讨青蒿琥酯(ART)通过介导铁死亡逆转结肠癌细胞对5-氟尿嘧啶(5-FU)耐药的作用。方法以人结肠癌HT-29/5-FU耐药细胞株为研究对象,采用MTS法检测5-FU、ART以及5-FU+ART对HT-29/5-FU细胞的抑制作用,并计算其逆转耐药倍数;克隆形成实验检测HT-29/5-FU细胞克隆形成能力;流式细胞术检测细胞内活性氧(ROS)水平;试剂盒检测细胞内丙二醛(MDA)水平;Western blotting检测细胞内Nrf2和GPX4蛋白表达水平。结果40μmol·L^(-1)ART可逆转HT-29/5-FU细胞对5-FU的耐药,逆转倍数为2.9倍;5-FU联合ART可抑制HT-29/5-FU细胞的克隆形成能力,升高细胞内ROS和MDA水平,并降低Nrf2及GPX4蛋白表达水平,且这些效应能够被铁死亡抑制剂Ferrostatin-1所逆转。结论ART能够通过抑制Nrf2、GPX4表达诱导铁死亡,从而逆转HT-29/5-FU细胞对5-FU的耐药。

Objective To investigate the effects of artesunate(ART)on the reverse of colon cancer cell resistance to 5-fluorouracil by mediating ferroptosis.Methods The inhibitory effects of 5-FU,ART and 5-FU+ART on HT-29/5-FU cells were detected by MTS,and the drug resistance index were calculated.The clone formation ability of HT-29/5-FU cells was detected by clone formation assay.The level of reactive oxygen species(ROS)was detected by flow cytometry.The level of malondialdehyde(MDA)was detected by MDA kit.The protein expression levels of Nrf2 and GPX4 were detected by West-ern blotting.Results 40μmol·L^(-1) ART could reverse the cell resistance to 5-FU,and the resistance index was 2.9.5-FU combined with ART could inhibit the clone formation ability of HT-29/5-FU cells,and increase the intracellular ROS and MDA levels,and decrease the protein expression levels of Nrf2 and GPX4.These effects could be reversed by ferroptosis in-hibitor ferrostatin-1.Conclusion ART could reverse the resistance of HT-29/5-FU cells to 5-FU by inhibiting the expres-sion of Nrf2 and GPx4 and inducing ferroptosis.