急性髓系白血病耐药细胞株的建立及Sirt1、PGC-1α的表达水平与耐药相关性的研究  

Establishment of Drug-Resistant Cell Lines of Acute Myeloid Leukemia and Correlation of Sirt1 and PGC-1α Expression Levels with Drug Resistance

作　　者：李历程 汪君娣 杨思思 周振 曾庆繁 郑方 LI Li-Cheng;WANG Jun-Di;YANG Si-Si;ZHOU Zhen;ZENG Qing-Fan;ZHENG Fang(Department of Hematology,Baiyun Hospital Affiliated to Guizhou Medical University,Guiyang 550014,Guizhou Province,China;Department of Pharmacy,Baiyun Hospital Affiliated to Guizhou Medical University,Guiyang 550014,Guizhou Province,China;Department of Anesthesiology,Baiyun Hospital Affiliated to Guizhou Medical University,Guiyang 550014,Guizhou Province,China)

机构地区：[1]贵州医科大学附属白云医院血液科,贵州贵阳550014 [2]贵州医科大学附属白云医院药剂科,贵州贵阳550014 [3]贵州医科大学附属白云医院麻醉科,贵州贵阳550014

出　　处：《中国实验血液学杂志》2022年第3期704-710,共7页Journal of Experimental Hematology

摘　　要：目的:构建耐阿糖胞苷的急性髓系白血病(AML)细胞株,探讨Sirt1、PGC-1α表达水平与耐药的相关性。方法:使用人急性原粒细胞白血病细胞Kasumi-1,采用Ara-C药物浓度逐渐增加的方法诱导细胞,通过CCK-8法检测加药前后Kasumi-1细胞的IC_(50)值,从而构建耐Ara-C细胞株;RT-qPCR检测Kasumi-1耐药株和亲本细胞株中Sirt1、PGC-1αmRNA的表达水平,蛋白免疫印迹Western blot检测Kasumi-1耐药细胞株和亲本细胞株中Sirt1、PGC-1α蛋白的表达水平。结果:构建后的Kasumi-1耐药细胞株具有常见的耐药细胞株的镜下形态特征,耐药指数大于5,表明Kasumi-1耐药株构建完成后具有良好的耐药性;RT-qPCR和Western blot检测发现在耐药细胞株中Sirt1和PGC-1αmRNA和蛋白的表达水平均高于亲本细胞株(P<0.001)。结论:采用浓度逐渐增加的方法可成功诱导出AML耐Ara-C细胞株,Sirt1与PGC-1α共同高表达可能介导了AML细胞耐药。Objective: To construct cytarabine-resistant acute myeloid leukemia(AML) cell lines, and explore the correlation between Sirt1, PGC-1α expression levels and drug resistance. Methods: Human acute promyelocytic leukemia Kasumi-1 cells were induced by the method of gradually increasing the concentration of Ara-C drug. The IC;value of Kasumi-1 cells before and after drug addition was detected by CCK-8 method, so as to construct Ara-C resistant cell lines. The expression levels of Sirt1 and PGC-1α mRNA in Kasumi-1 drug-resistant cell lines and their parental cell lines were detected by real-time fluorescence quantitative PCR, and the expression levels of Sirt1 and PGC-1α protein in kasumi-1 drug-resistant cell lines and their parental cell lines were detected by Western blot. Results: The constructed Kasumi-1 cell line had common morphological characteristics of drug-resistant cell lines under microscope, and the drug resistance index was greater than 5, indicating that Kasumi-1 drug-resistant cells had good drug resistance after the construction. The RT-qPCR and Western blot assays showed that the expression levels of Sirt1 and PGC-1α mRNA and protein in the drug-resistant cell lines were higher than those of the parental cell lines(P<0.001). Conclusion: AML cell lines resistant to Ara-C can be successfully induced by the method of gradually increasing the concentration, and the co-high expression of Sirt1 and PGC-1α may mediate the drug resistance of AML cells.

关 键 词：急性髓细胞白血病 细胞耐药 SIRT1 PGC-1Α 

分 类 号：R733.71[医药卫生—肿瘤]