急性淋巴细胞白血病耐药细胞株的建立及耐药机制的研究  

Establishment of Drug-resistant Acute Lymphoblastic Leukemic Cell Lines and Their Resistance Mechanism

作　　者：李田田 张莉[1,2] 陶善东[1,2] 马晶晶[1,2] 于亮 LI Tian-Tian;ZHANG Li;TAO Shan-Dong;MA Jing-Jing;YU Liang(Department of Hematology, Huai＇an First People＇s Hospital Affiliated to Nanjing Medical University, Huai＇an 223300, Jiangsu Province, China;Key Laboratory of Hematology, Nanjing Medical University, Nanjing 210029, Jiangsu Province, China)

机构地区：[1]南京医科大学附属淮安第一医院血液科,江苏淮安223300 [2]南京医科大学血液病重点实验室,江苏南京210029

出　　处：《中国实验血液学杂志》2018年第3期698-704,共7页Journal of Experimental Hematology

基　　金：江苏省自然科学基金资助项目(BK20141254);淮安市"533英才工程"基金资助项目(HAA201739)

摘　　要：目的:构建耐阿霉素的急性淋巴细胞白血病(ALL)细胞株,并探讨其耐药机制。方法:采用阿霉素(ADR)小剂量浓度递增间歇诱导SUP-B15(Ph^+)和RS4;11(Ph^-)细胞,建立SUP-B15/ADR和RS4;11/ADR耐药细胞株;应用CCK-8法检测细胞的活力与增殖;Western blot检测P-糖蛋白(P-gp)的表达;RT-PCR检测MDR1基因的表达。结果:成功构建ALL耐药细胞株SUP-B15/ADR和RS4;11/ADR,耐药指数分别为14.088±0.763和10.473±1.024。冻存的SUP-B15/ADR和RS4;11/ADR细胞复苏后,细胞存活率分别为88.4±1.2%和89.3±1.6%,耐药指数分别为13.976±0.967和10.342±0.846,与冻存前相比差异无统计学意义(P﹥0.05)。撤药培养1个月,2株细胞的耐药指数明显降低,分别为12.893±1.255和9.327±0.321(P﹤0.05)。SUP-B15/ADR和RS4;11/ADR细胞对阿糖胞苷的耐药指数分别为3.459±0.451和3.992±0.795,对依托泊苷的耐药指数分别为3.564±0.365和3.654±0.251。MDR1和P-gp在耐药细胞中的表达较亲本细胞增高。结论:小剂量浓度递增间歇诱导法可成功诱导出ALL耐阿霉素细胞株,其耐药机制是上调MDR1与P-gp的表达。Objective： To establish the adriamycin（ ADR）-resistant ALL cell lines and to investigate their drug-resistan mechanisms.Methods： The drug-resistant cell lines SUP-B15/ADR and RS4; 11/ADR were derived by exposing the parental cells[SUP-B15（ Ph-＋） and RS4;11（ Ph--）] to the ascending concentrations of ADR.The cell viability was detected by CCK-8 method.The expression of P-gp was examined by Western blot,and RT-qPCR was performed to detect the expression of MDR1.Results： The drug-resistant cell lines SUP-B15/ADR and RS4;11/ADR were successfully established,their resistance indexes were 14.088 ± 0.763 and 10.473 ± 1.024,respectively.After the cryopreserved SUP-B15/ADR and RS4;11/ADR cells were resuscitated,their survival rates were 88.4 ± 1.2% and 89.3 ± 1.6% respectively,while their resistance indexes were 13.976 ± 0.967 and 10.342 ± 0.846 respectively（ P〉0.05）.When the drug-resistant cells were cultured in the medium without ADR for 1 month,their drug-resistance indexes dropped down to 12.893 ± 1.255 and 9.327 ± 0.321 respectively（ P〈0.05）.Drug-resistant cell lines had the cross-resistance to cytarabine and etoposide.The expression of P-gp and MDR1 in drug-resistant cells was significantly higher than that in wild-type cells.Conclusion： Two drug-resistant ALL cell lines have been successfully established by exposing to the ascending concentration of ADR.The over-expression of MDR1 and P-gp in drug-resistant cells may be one of the mechanisms underlying the drug resistance.

关 键 词：急性淋巴细胞白血病 耐药细胞株 阿霉素 P-糖蛋白 MDR1 

分 类 号：R733.71[医药卫生—肿瘤]