PI3Kδ抑制剂idelalisib对人髓系白血病细胞增殖的抑制作用及对阿霉素耐药性的逆转作用  

Inhibitory effect of PI3Kδinhibitor idelalisib on proliferation of human myeloid leukemia cells and the reversal effect on drug resistance to adriamycin

作　　者：李昆仑 易平勇 罗晗嘉 李基炜 孟柳 唐敏 曾维斯 杨烁 王伟 LI Kunlun;YI Pingyong;LUO Hanjia;LI Jiwei;MENG Liu;TANG Min;ZENG Weisi;YANG Shuo;WANG Wei(Department of Geriatrics/Department of Gastroenterology and Urology WardⅡ,Hunan Cancer Hospital/Cancer Hospital Affiliated to Xiangya School of Medicine,Central South University,Changsha 410013,China)

机构地区：[1]中南大学湘雅医学院附属肿瘤医院,湖南省肿瘤医院老年特需/消化泌尿内二科,长沙410013

出　　处：《中南大学学报（医学版）》2020年第12期1389-1397,共9页Journal of Central South University ：Medical Science

基　　金：湖南省科技厅重点课题(2014TT1001);湖南省卫生健康委员会重点课题(20201665);中国临床肿瘤学会课题(Y-HR2018-234)。

摘　　要：目的:观察阿霉素(adriamycin,ADM)单药、艾德拉尼(idelalisib)单药以及ADM联合idelalisib对人髓系白血病细胞株K562和K562/ADM的增殖以及细胞内ADM相对浓度的影响,探讨idelalisib对K562和K562/ADM两种细胞株ADM耐药的逆转作用及其机制。方法:以K562及K562/ADM细胞株为观察对象,分别以不同浓度ADM及idelalisib进行干预,计算ADM对两种细胞株的半抑制浓度(50%inhibitory concentration,IC50)及耐药指数(resistance index,RI);计算idelalisib对于两种细胞株的无细胞毒性作用浓度(细胞抑制率<5%)。后续将K562和K562/ADM细胞株分别分为ADM组和ADM+idelalisib组。分别用MTT法、生物发光法和流式细胞术检测各组细胞存活率、细胞株内ATP和ADM水平的变化。结果:随着ADM浓度的增加,K562及K562/ADM细胞存活率相应降低,呈剂量依赖性;ADM在K562与K562/ADM细胞株中的IC50分别为0.2 mg/L和1.0 mg/L,RI为5。随着idelalisib浓度的增加,K562及K562/ADM细胞存活率也相应降低,呈剂量依赖性;idelalisib对K562和K562/ADM细胞株的无细胞毒性体积浓度(抑制率<5%)分别约为25μmol/L和15μmol/L。MTT法结果显示:在K562/ADM细胞株中,ADM+idelalisib组的细胞存活率低于ADM组,差异有统计学意义(P<0.05);在K562细胞株中,ADM+idelalisib组的细胞存活率与ADM组相比差异无统计学意义(P>0.05)。生物发光法结果显示:K562细胞株内ATP水平为(91.502±0.479)mmol/L,K562/ADM细胞株内ATP水平为(24.311±0.349)mmol/L。在K562/ADM细胞株中,ADM+idelalisib组ATP水平低于ADM组,差异有统计学意义(P<0.05);在K562细胞株中,ADM+idelalisib组ATP水平与ADM组相比差异无统计学意义(P>0.05)。流式细胞术检测结果显示:在K562/ADM细胞株中,ADM+idelalisib组ADM吸收率高于ADM组,差异有统计学意义(P<0.05);在K562细胞株中,ADM+idelalisib组ADM吸收率与ADM组相比差异无统计学意义(P>0.05)。在K562/ADM细胞株中,ADM+idelalisib组ADM排除率低于ADM组,差异有统计学意义(Objective:To investigate the effect of adriamycin(ADM),idelalisib or ADM and their combination on cell proliferation and intracellular concentration of ADM,and to explore the reversal effect of idelalisib on drug resistance to ADM.Methods:The K562 and K562/ADM cells were respectively treated with ADM and idelalisib at different concentrations.The 50%inhibitory concentration(IC50)and drug resistance index(RI)of ADM to the 2 kinds of cells were measured by methyl thiazolyl tetrazolium(MTT)assay.Non-cytotoxic dose(cell inhibition rate<5%)of idelalisib in the 2 kinds of cells was determined.Then the K562 and k562/ADM cells were divided into the following groups:a K562 cells+ADM group,a K562 cells+ADM+idelalisib group,a K562/ADM cells+ADM group,and a K562/ADM cells+ADM+idelalisib group.The survival rates,the intracellular ATP levels,and the relative concentration of intracellular ADM were detected by MTT method,ATP bioluminescence assay(ATP-BLA)and flow cytometry(FCM),respectively.Results:The cell survival rates were significantly decreased in a dose-dependent manner when the cells were treated with different doses of ADM(0.001-10.000 mg/L).The IC50 value of ADM in the K562 and K562/ADM cells were 0.2 mg/L and 1.0 mg/L,respectively.The RI value was 5.The cell survival rates were also significantly decreased in a dose-dependent manner when the cells were treated with different doses of idelalisib(1-50μmol/L).The non-cytotoxic dose of idelalisib in the K562 and K562/ADM cells were 25μmol/L and 15μmol/L,respectively.The cell survival rates in the ADM+idelalisib group was less than that in the ADM group(P<0.05);while there was no statistical difference between the ADM+idelalisib group and the ADM group in the K562 cells(P>0.05).The intracellular ATP level in the K562 cells was about(91.502±0.479)mmol/L,and that in the K562/ADM cells was about(24.311±0.349)mmol/L.The intracellular ATP level in the ADM+idelalisib group in the K562/ADM cells was less than that in the ADM group(P<0.05);but there was no statistical differ

关 键 词：idelalisib 白血病 K562细胞 K562/ADM细胞 肿瘤多发耐药 磷脂酰肌醇3-激酶/蛋白激酶B信号通路 逆转耐药 

分 类 号：R733.7[医药卫生—肿瘤]