阿伐他汀通过PTEN/mTOR信号调节糖酵解代谢逆转白血病耐药的作用及机制  

Effect and Mechanism of Atorvastatin on Reversing Drug Resistance in Leukemia by Regulating Glycolysis through PTEN/mTOR Pathway

作　　者：刘苗[1] 周攀 LIU Miao;ZHOU Pan(Department of Paediatrics,Renmin Hospital of Wuhan University,Wuhan 430060,Hubei Province,China;Hubei Medical Devices Quality Supervision and Test Institute,Wuhan 430075,Hubei Province,China)

机构地区：[1]武汉大学人民医院儿科,湖北武汉430060 [2]湖北省医疗器械质量监督检验研究院,湖北武汉430075

出　　处：《中国实验血液学杂志》2023年第1期38-44,共7页Journal of Experimental Hematology

基　　金：湖北省自然科学基金项目(2020CFB611)。

摘　　要：目的:探讨阿伐他汀对耐阿霉素人急性早幼粒白血病细胞系HL-60/ADM糖酵解代谢的影响及作用机制。方法:取对数生长期耐阿霉素白血病细胞HL-60/ADM,给予不同浓度阿伐他汀处理后,采用CCK-8法测定细胞增殖活性,流式细胞术检测细胞凋亡,葡萄糖消耗实验检测白血病细胞糖酵解活性,Western blot方法检测PTEN、p-m TOR、PKM2、HK2、P-gp、MRP1蛋白的表达;将PTEN-si RNA转染至HL-60/ADM细胞后,进一步采用上述方法检测PTEN低表达对阿伐他汀调节HL-60/ADM细胞凋亡及糖酵解代谢的影响。结果:CCK-8结果显示,阿伐他汀呈浓度依赖性和时间依赖性抑制HL-60/ADM细胞增殖(r=0.872,r=0.936),10μmol/L阿伐他汀干预24 h后HL-60/ADM细胞增殖活性下降最明显,增殖活性降至(32.3±2.18)%。流式细胞术结果显示,阿伐他汀诱导HL-60/ADM细胞凋亡,该作用呈浓度依赖性(r=0.796),10μmol/L阿伐他汀对HL-60/ADM细胞的诱导凋亡作用最强,细胞凋亡率达到(48.78±2.95)%。葡萄糖消耗实验结果表明阿伐他汀明显抑制HL-60/ADM细胞糖酵解活性,该作用呈浓度依赖性和时间依赖性(r=0.915,r=0.748),10μmol/L阿伐他汀干预24 h后对糖酵解活性的抑制作用最强,相对葡萄糖消耗量降至(46.53±1.71)%。Western blot结果显示,给予阿伐他汀干预24 h后,p-m TOR、PKM2、HK2、P-gp、MRP1蛋白表达呈浓度依赖性降低(r=0.737,r=0.695,r=0.829,r=0.781,r=0.632),而PTEN蛋白表达呈浓度依赖性升高(r=0.531)。进一步将PTEN-si RNA转染HL-60/ADM细胞,结果显示,低表达PTEN减弱了阿伐他汀对细胞凋亡的促进作用以及对糖酵解代谢和多药耐药的抑制作用。结论:阿伐他汀能抑制白血病耐药株HL-60/ADM细胞增殖,诱导细胞凋亡,抑制糖酵解代谢,推测机制可能是通过上调PTEN表达,抑制m TOR激活,下调PKM2和HK2糖酵解代谢基因表达,从而逆转耐药。Objective: To investigate the influence and mechanism of atorvastatin on glycolysis of adriamycin resistant acute promyelocytic leukemia(APL) cell line HL-60/ADM. Methods: HL-60/ADM cells in logarithmic growth phase were treated with different concentrations of atorvastatin, then the cell proliferation activity was measured by CCK-8 assay, the apoptosis was detected by flow cytometry, the glycolytic activity was checked by glucose consumption test, and the protein expressions of PTEN, p-m TOR, PKM2, HK2, P-gp and MRP1 were detected by Western blot. After transfection of PTEN-si RNA into HL-60/ADM cells, the effects of low expression of PTEN on atorvastatin regulating the behaviors of apoptosis and glycolytic metabolism in HL-60/ADM cells were further detected. Results: CCK-8 results showed that atorvastatin could inhibit the proliferation of HL-60/ADM cells in a concentration-dependent and timedependent manner(r=0.872, r=0.936), and the proliferation activity was inhibited most significantly when treated with 10 μmol/L atorvastatin for 24 h, which was decreased to(32.3±2.18)%. Flow cytometry results showed that atorvastatin induced the apoptosis of HL-60/ADM cells in a concentration-dependent manner(r=0.796), and the apoptosis was induced most notably when treated with 10 μmol/L atorvastatin for 24 h, which reached to(48.78±2.95)%. The results of glucose consumption test showed that atorvastatin significantly inhibited the glycolytic activity of HL-60/ADM cells in a concentration-dependent and time-dependent manner(r=0.915, r=0.748), and this inhibition was most strikingly when treated with 10 μmol/L atorvastatin for 24 h, reducing the relative glucose consumption to(46.53±1.71)%. Western blot indicated that the expressions of p-m TOR, PKM2, HK2, P-gp and MRP1 protein were decreased in a concentrationdependent manner(r=0.737, r=0.695, r=0.829, r=0.781, r=0.632), while the expression of PTEN protein was increased in a concentration-dependent manner(r=0.531), when treated with different concentrations of ator

关 键 词：阿伐他汀 白血病耐药 PTEN/mTOR通路 糖酵解 

分 类 号：R733.7[医药卫生—肿瘤]