Objective: To investigate the effect and involved mechanism of RSL3 on ferroptosis action in acute leukemia cells MOLM13 and its drug-resistant cells.
Methods: After MOLM13 treated with RSL3, CCK-8 assay was performed to detect cell viability, flow cytometry was used to detect the reactive oxygen species (ROS) level of the cells, RT-qPCR and Western blot were used to detect the expression of glutathione peroxidase 4 (GPX4). After MOLM13/IDA and MOLM13/Ara-C, the drug-resistant cell lines were constructed, the ferroptosis induced by RSL3 was observed. Bone marrow samples were collected from patients with acute monocytic leukemia. RT-qPCR and Western blot were performed to detect the expression of related genes and proteins involved in ferroptosis pathway.
Results: RSL3 significantly inhibited the cell viability of MOLM13 and increased the intracellular ROS level, which were partially reversed by ferrostatin-1. The mRNA and protein expression of GPX4 decreased in MOLM13 treated with RSL3. RSL3 inhibited the viability of MOLM13/IDA and MOLM13/Ara-C cells more strongly than that of non-drug resistant cells, also increased the intracellular ROS level . The cytotoxic effects were partially reversed by ferrostatin-1. The mRNA and protein expressions of GPX4 in MOLM13/IDA and MOLM13/Ara-C cells were higher than those in non-drug resistant cells. The mRNA and protein levels of GPX4 in bone marrow of relapsed/refractory acute mononuclear leukemia patients were higher than those of ordinary acute mononuclear leukemia patients.
Conclusion: RSL3 can induce non-drug resistant cells MOLM13 ferroptosis by inhibiting GPX4 activity. MOLM13/IDA and MOLM13/Ara-C are more sensitive to RSL3 compared with non-drug resistant cells MOLM13, which may be caused by the differences in GPX4 expression. The expressions of GPX4 mRNA and protein in relapsed/refractory acute mononuclear leukemia are higher than those in ordinary acute mononuclear leukemia.
题目: RSL3诱导急性白血病细胞株MOLM13及其耐药细胞株发生铁死亡的作用及相关机制研究.
目的: 探讨铁死亡激活剂RSL3诱导急性白血病细胞株MOLM13及其耐药细胞株发生铁死亡的作用及相关机制。.
方法: 急性白血病细胞株MOLM13与RSL3共同培养，应用CCK-8法检测细胞的增殖活性，应用流式细胞术检测细胞内活性氧（ROS）水平的变化，应用RT-qPCR、Western blot技术检测细胞内谷胱甘肽过氧化物酶4（GPX4）的表达。构建MOLM13/IDA、MOLM13/Ara-C耐药细胞株，检测RSL3诱导耐药肿瘤细胞发生铁死亡的现象。收集急性单核细胞白血病患者骨髓样本，应用RT-qPCR、Western blot技术分别检测样本中铁死亡通路相关基因及蛋白的表达。.
结果: RSL3对MOLM13细胞增殖活性具有明显抑制作用，细胞内的ROS水平增高，铁死亡特异性抑制剂ferrostatin-1可以部分逆转这种现象，经RSL3作用后MOLM13细胞内GPX4基因及蛋白表达下降。RSL3对MOLM13/IDA、MOLM13/Ara-C细胞生长抑制作用较非耐药细胞增强，细胞内的ROS水平增高，ferrostatin-1可以部分逆转这种现象。MOLM13/IDA、MOLM13/Ara-C细胞内GPX4基因及蛋白表达较非耐药细胞增高。复发、难治性急性单核白血病患者骨髓样本中GPX4基因及蛋白表达相较于普通急性单核白血病患者骨髓样本表达增高。.
结论: RSL3通过抑制GPX4活性诱导非耐药细胞株MOLM13发生铁死亡。MOLM13/IDA、MOLM13/Ara-C较非耐药细胞对RSL3的抑制作用更加敏感，可能基于细胞内GPX4表达差异。复发、难治性急性单核细胞白血病中GPX4基因及蛋白表达高于普通急性单核细胞白血病。.