米哚妥林衍生物L156对人红白血病HEL细胞增殖和凋亡的作用及其机制研究  

Effects and Mechanism of Midoletolin Derivative L156 on HEL Cells of Human Erythrohematosis

作　　者：王春林 龙群[1] 王立平[1] 朱伟明 王昌西[3] 李艳梅 WANG Chunlin;LONG Qun;WANG Liping;ZHU Weiming;WANG Changxi;LI Yanmei(The Key Laboratory of Chemistry for Natural Products of Guizhou Province and Chinese Academy of Sciences,Guiyang,Guizhou,550014,China;Ocean University of China,Qingdao,Shandong,266100,China;The People's Hospital of Anshun City,Anshun,Guizhou,561000,China)

机构地区：[1]贵州省中国科学院天然产物化学重点实验室,贵州贵阳550014 [2]中国海洋大学,山东青岛266100 [3]安顺市人民医院,贵州安顺561000

出　　处：《肿瘤药学》2020年第1期66-72,共7页Anti-Tumor Pharmacy

基　　金：贵州省科技支撑项目重点项目(QKH20192762);贵州省科学基础研究重点项目(QHK20181409);贵州省科技计划项目(黔科合LH字[2016]7412)

摘　　要：目的研究米哚妥林衍生物L156对人红白血病HEL细胞增殖和凋亡的影响及其作用机制。方法采用MTT法检测L156对人红白血病HEL细胞增殖的影响,绘制细胞生长曲线并计算IC50;流式细胞仪检测L156不同浓度和作用时间对HEL细胞分化、周期和凋亡的影响;Hoechst 33258染色检测L156对细胞核形态的影响;Western blotting检测L156对HEL细胞中凋亡和周期相关蛋白的影响。结果L156可显著抑制人红白血病HEL细胞的增殖,IC50为(0.073±0.0034)μM;L156可诱导HEL细胞发生早期凋亡和晚期凋亡,并呈时间和浓度依赖性(P<0.05);L156可阻滞HEL细胞周期,使G2/M期细胞比例显著增加;L156可明显提高细胞巨核分化标志物CD41a/CD61和细胞红系分化物CD71/CD235a的表达水平,并呈浓度依赖性(P<0.05);L156可抑制细胞信号转录因子STAT3的磷酸化,下调抗凋亡蛋白Bcl-2和周期蛋白cyclinB1的表达,上调凋亡蛋白Cleaved-Caspase-3的表达,差异均有统计学意义(P<0.05)。结论L156可抑制STAT3的磷酸化,从而通过下调Bcl-2的表达和上调Cleaved-Caspase-3的表达来促进HEL细胞凋亡,并通过下调周期蛋白cyclinB1的表达将细胞周期阻滞于G2/M期,从而抑制细胞增殖,但其靶点是否为STAT3仍需进一步研究确定。L156还可诱导HEL细胞向巨核和红系两个方向分化,使其具有向良性转变的可能,可作为治疗红白血病的候选药物。Objective To investigate the effects of midostaurin derivative Ll56 on the activity of human erythroleukemia(HEL)cells and the possible mechanism.Methods MTT assay was used to detect the effects of L156 on the proliferation of HEL cells.And the IC50 value was calculated and growth inhibition curve was made.Flow cytometry was used to determine the effects of different concentrations of L156 and different treating time on the differentiation,cycle and apoptosis of HEL cells.The changes of cell morphology of HEL cells were detected by Hoechst 33258 staining.Western blotting was applied to detect the effects of L156 on the apoptosis-and cycles-related proteins in HEL cells.Results L156 had a significant inhibitory effect on the proliferation of HEL cells,and the IC50 value was(0.073±0.0034)μM.L156 also induced both early and late apoptosis of HEL cells in a time-and concentration-dependent manner(P<0.05).It blocked the cell cycles and increased the cell ratio in G2/M phase.Moreover,the expression of megakaryocyte differentiation marker CD41 a/CD61 and erythrocyte differentiation marker CD71/CD235 a were significantly increased in a concentration-dependent way(P<0.05).Besides,L156 retrained the phosphorylation of cell signal transcription factor 3(STAT3),down-regulated the expressions of protein Bcl-2 and cyclinB1,and upregulated the expression of Cleaved-Caspase-3(P<0.05).Conclusion L156 restrained the phosphorylated STAT3(p-STAT3),reduced the expression of Bcl-2 in HEL cells and promoted the formation of Cleaved-Caspase-3 to achieve cell apoptosis.It inhibited the proliferation of HEL cells by down-regulating the expression of cyclinB 1 and blocking the G2/M phase of cell cycle.However,further studies are needed to determine whether its target is STAT3.In addition,L156 could induce HEL cells to differentiate into megakaryocyte and erythrocyte lines,making it possible for them to make a benign transition.L156 could senve as a candidate drug for the treatment of erythroleukemia.

关 键 词：白血病 米哚妥林衍生物 L156 凋亡 细胞周期 STAT3 

分 类 号：R733.7[医药卫生—肿瘤]