自噬水平与白血病K562/ADM细胞耐药的关系研究  

Study on relationship between autophagy and drug resistance of leukemia K562/ADM cells

作　　者：李彩丽[1] 高静[2] 黄双盛[1] 赵永勋[3] 刘进 黄涛[1] 李敏[1] LI Cai-li;GAO Jing;HUANG Shuang-sheng;ZHAO Yong-xun;LIU Jin;HUANG Tao;LI Min(Dept of Biochemistry,School of Medicine of Northwest Minzu University,Lanzhou 730030,China;Affiliated Hospital of Northwest Minzu University,Lanzhou 730030,China;Dept of Surgical Oncology,the First Affiliated Hospital of Lanzhou University,Lanzhou 730000,China)

机构地区：[1]西北民族大学医学部生物化学教研室 [2]西北民族大学附属医院检验科,甘肃兰州730030 [3]兰州大学第一附属医院肿瘤外科,甘肃兰州730000

出　　处：《中国药理学通报》2021年第3期343-348,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81660028);西北民族大学引进人才项目(No 31920170038);甘肃省科技计划项目(No 17JR5RA272);西北民族大学科研创新团队。

摘　　要：目的采用阿霉素(adriamycin,ADM)“逐步提高药物浓度+间歇性诱导”的方式体外诱导建立稳定耐受15μmol·L^(-1) ADM的白血病K562/ADM细胞,观察该细胞对其它化疗药物的敏感性以及细胞自噬水平与耐药的关系。方法MTT法检测细胞对几种化疗药物的敏感性;用透射电镜、荧光显微镜观察细胞自噬形态学改变;Annexin-V/PI双染流式细胞仪检测细胞凋亡;Western blot检测自噬和耐药相关蛋白的表达水平。结果K562/ADM细胞除了对ADM产生明显耐药外,还对多种化疗药物如:吡柔比星、柔红霉素、5-FU和长春新碱等有交叉耐药,但对三氧化二砷较敏感。K562/ADM细胞内自噬体数量、MDC荧光强度以及LC3Ⅰ/Ⅱ、Beclin-1蛋白表达水平均高于亲本细胞。用3-MA抑制自噬可明显增加K562/ADM细胞对ADM的敏感性,同时也能有效抑制K562/ADM细胞内耐药相关蛋白的表达。结论K562/ADM细胞出现多药耐药现象,且耐药性与细胞自噬水平有密切关系。Aim To set up leukemic K562/ADM cells with stable tolerance to 15μmol·L^(-1) ADM induced in vitro by long-term and continuous stepwise increment of adriamycin(ADM)concentration,to observe the sensitivity to other chemotherapy drugs and the relationship between autophagy and drug resistance.Methods MTT assay was used to detect the sensitivity of cells to chemotherapy drugs.The morphological changes of autophagy were observed by transmission electron microscope and fluorescence microscopy.Cell apoptosis analysis was performed using Annexin-V/PI double staining and flow cytometry(FCM).The expressions of autophagy and drug resistance associated proteins were tested by Western blot.Results K562/ADM cells were cross-resistance to the other chemotherapeutics besides adriamycin,such as pirarubicin,daunorubicin,5-flurouracil,vincristine but not arsenic trioxide.The number of autophagosomes,the fluorescence intensity of monodansylcadaverine(MDC)and the expression of LC3-Ⅱ,Beclin-1 in K562/ADM cells were significantly higher than those in K562 cells.The inhibition of autophagy by 3-MA significantly increased the sensitivity of K562/ADM cells to ADM,and 3-MA also effectively inhibited the expressions of drug resistance related proteins P-gp,MRP1 and BCRP in K562/ADM cells.Conclusions The K562/ADM cells resistant to adriamycin occur multidrug resistance,and the drug resistanceis closely related to the level of autophagy.

关 键 词：多药耐药 自噬 阿霉素 K562细胞 K562/ADM细胞 白血病 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R733.7[医药卫生—基础医学]