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

To construct cytarabine-resistant acute myeloid leukemia (AML) cell lines, and explore the correlation between Sirt1, PGC-1α expression levels and drug resistance.Human acute promyelocytic leukemia Kasumi-1 cells were induced by the method of gradually increasing the concentration of Ara-C drug. The IC50 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.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).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.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.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.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IC50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-1α mRNA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.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-1α mRNA和蛋白的表达水平均高于亲本细胞株（P<0.001）.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.