摘要
目的:构建稳定的耐伊马替尼(IM)的费城染色体阳性(Ph^+)急性B淋巴细胞白血病(B-ALL)细胞株(SUP-B15/IM),为其耐药机制研究提供细胞模型。方法:采用IM小剂量浓度递增间歇诱导亲本细胞(SUP-B15),建立对IM耐药的SUP-B15细胞株(耐药组,SUP-B15/IM)模型,并用与耐药组细胞培养条件相同但不加IM的SUP-B15作为对照细胞(对照组,SUP-B15/C),采用CCK-8法检测SUP-B15、SUP-B15/C及SUP-B15/IM对IM的耐药性,用流式细胞术检测细胞凋亡率,用RT-PCR检测各组细胞相关耐药基因MDR 1和MRP 5的表达。结果:成功建立SUP-B15/IM耐药细胞株,其对IM的耐药指数为12.3;用100μmol/L IM刺激细胞24 h后,SUP-B15/C的凋亡率远大于SUP-B15/IM细胞(P<0.001);100μmol/L IM浓度下,SUP-B15/IM细胞中MDR 1和MRP 5 mRNA表达水平显著高于SUP-B15/C细胞,差异有统计学意义(P<0.001)。结论:成功构建了耐IM的Ph^+B-ALL细胞株SUP-B15/IM。
Objective:To construct a stable imatinib-resistant(IM)philadelphia chromosome-positive(Ph^+)acute B lymphoblastic leukemia(B-ALL)cell line(SUP-B15/IM)to provide a cell model for the study of its resistance mechanism.Methods:Cell lines of IM resistant SUP-B15(drug resistance group,SUP-B15/IM)were established to induce parent cells(SUP-B15)with increasing dose concentration.The SUP-B15,SUP-B15/C and SUP-B15/IM were detected by CCK-8 method using the same SUP-B15 without IM as the cell culture conditions of drug-resistant group(control group,SUP-B15/C)Drug resistance to IM.The apoptosis rate was detected by flow cytometry,and the expression of cell-related drug resistance gene MDR1 and MRP5 was detected by RT-PCR.Results:The resistance index of SUP-B15/IM resistant cell line to IM was 12.3.The apoptosis rate of SUP-B15/C was much higher than that of SUP-B15/IM cells after 24 h stimulation with 100μmol/L IM cells(P<0.001).The expression levels of MDR1 and MRP5 mRNA in SUP-B15/IM cells were significantly higher than those in SUP-B15/C cells at 100μmol/L IM concentration,the difference was statistically significant(P<0.001).Conclusion:The SUP-B15/IM of IM resistant Ph^+B-ALL cell lines is successfully constructed.
作者
方宗宇
马莉
张智琪
吴青青
FANG Zongyu;MA Li;ZHANG Zhiqi;WU Qingqing(The Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine,Guizhou Medical University,Guiyang 550004,Guizhou,China;Department of Clinical Laboratory,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China)
出处
《贵州医科大学学报》
CAS
2020年第4期415-421,共7页
Journal of Guizhou Medical University
基金
国家自然科学基金(31460236)。
关键词
白血病
费城染色体
伊马替尼
耐药
leukemia
philadelphia chromosome
imatinib
drug resistance
