Ponatinib对人慢性髓系白血病细胞株K562自噬的影响  

Effect of Ponatinibon Autophagy of Chronic Myeloid Leukemia CellLine K562

作　　者：张蕾[1] 吴一凡 郭圣洁 孙青颖 赵志明 陈薇潼 徐小冬[1] 赵冬久[1] 朴正浩 ZHANG Lei;WU Yifan;GUO Shengjie;SUN Qingying;ZHAO Zhiming;CHEN Weitong;XU Xiaodong;ZHAO Dongjiu;PIAO Zhenghao(School of Basic Medical Sciences,Hangzhou Normal University,Hangzhou 311121,China)

机构地区：[1]杭州师范大学基础医学院,浙江杭州311121

出　　处：《杭州师范大学学报（自然科学版）》2024年第4期395-401,共7页Journal of Hangzhou Normal University(Natural Science Edition)

基　　金：浙江省自然科学基金项目(LY23H010001);杭州师范大学“本科生创新能力提升工程”项目;杭州师范大学2023年虚拟教研室建设项目(4255C5052300115).

摘　　要：酪氨酸激酶抑制剂(tyrosine kinase inhibitors,TKIs)是治疗白血病的靶向药物,其中第3代的Ponatinib疗效最强、见效最快,但与其他TKIs在作用机制上存在的差异尚不明确.为此,采用第1代的Imatinib和第3代的Ponatinib处理慢性髓系白血病细胞株K562细胞,观察活细胞数量、线粒体活性、活性氧(reactive oxygen species,ROS)和自噬相关分子的变化.经研究发现,Ponatinib比Imatinib更显著地抑制K562细胞的增殖和生存率,并降低了线粒体活性,但ROS没有明显差异;免疫印迹实验发现Ponatinib比Imatinib更明显促进自噬标志性分子LC3B蛋白的表达,并显著降低自噬受体蛋白p62的表达,但未检测到AMPKα、ULK1和Beclin1蛋白的磷酸化.此外,Bafilomycin A1能阻止Ponatinib对p62蛋白的抑制作用,而Brefeldin A和N-乙酰-L-半胱氨酸未能起到阻止作用.这些结果表明,Ponatinib比Imatinib更显著地引起K562细胞的线粒体损伤,进而诱导更明显的自噬,这一发现可能为解释Ponatinib疗效更强的作用机制提供重要线索.Tyrosine kinase inhibitors(TKIs)are targeted drugs for the treatment of leukemia.At present,there are three generations of them,among which the third-generation Ponatinib has the strongest effect and the fastest response,but the difference in the mechanism between other TKIs is not clear.To investigate the differences,chronic myeloid leukemia K562 cells were treated with first-generation Imatinib and third-generation Ponatinib.The changes in the number of viable cells,mitochondrial activity,reactive oxygen species(ROS)and autophagy related molecules were observed.Ponatinib inhibited the proliferation and survival rate of K562 cells more significantly than Imatinib,and reduced mitochondrial activity,but there was no significant difference in ROS.Compared with Imatinib,Ponatinib significantly promoted the autophagy marker LC3B protein and reduced the autophagy receptor protein p62,while the phosphorylation of AMPKα,ULK1 and Beclin1 proteins was not detected.In addition,Bafilomycin A1,but not Brefeldin A and N-acetyl-L-cysteine,blocked the inhibitory effect of Ponatinib on p62 protein.These results suggest that Ponatinib caused mitochondrial damage in K562 cells more significantly than Imatinib,which in turn induced more pronounced autophagy,a finding that may provide important clues to explain the stronger therapeutic effect of Ponatinib.

关 键 词：慢性髓系白血病 PONATINIB 酪氨酸激酶抑制剂 线粒体 自噬 

分 类 号：Q291[生物学—细胞生物学]