贵州医科大学学报

2020, v.45;No.235(04) 415-421

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Ph~+急性B淋巴细胞白血病耐伊马替尼细胞模型的建立
Establishment of Imatinib Cell Model of Ph~+ Acute B Lymphoblastic Leukemia

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;Department of Clinical Laboratory,the Affiliated Hospital of Guizhou Medical University;

摘要(Abstract):

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(SUPB15),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MDR1和MRP5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MDR1和MRP5 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 chromosomepositive( 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 100 μmol/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 100 μmol/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.

关键词(KeyWords): 白血病;费城染色体;伊马替尼;耐药
leukemia;philadelphia chromosome;imatinib;drug resistance

Abstract:

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基金项目(Foundation): 国家自然科学基金(31460236)

作者(Author): 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;Department of Clinical Laboratory,the Affiliated Hospital of Guizhou Medical University;

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DOI: 10.19367/j.cnki.1000-2707.2020.04.008

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