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  

Effect and Involved Mechanism of RSL3-induced Ferroptosis in Acute Leukemia Cells MOLM13 and Drug-resistant Cell Lines

作　　者：程霖 金鑫 卢文艺[3] 孙瑞 曹雅青 魏运雄 王露桥 贺小圆 袁婷 孟娟霞[3] 赵明峰[3] CHENG Lin;JIN Xin;LU Wen-Yi;SUN Rui;CAO Ya-Qing;WEI Yun-Xiong;WANG Lu-Qiao;HE Xiao-Yuan;YUAN Ting;MENG Juan-Xia;ZHAO Ming-Feng(Department of Hematology,The First Central Clinical College of Tianjin Medical University;Nankai University School of Medicine;Department of Hematology,Tianjin First Central Hospital,Tianjin 300192,China)

机构地区：[1]天津医科大学第一中心临床学院血液科 [2]南开大学医学院 [3]天津市第一中心医院血液科,天津300192

出　　处：《中国实验血液学杂志》2021年第4期1109-1118,共10页Journal of Experimental Hematology

摘　　要：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。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 sensitiveto 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.

关 键 词：急性白血病 耐药细胞 铁死亡 谷胱甘肽过氧化物酶4 活性氧 

分 类 号：R733.71[医药卫生—肿瘤]