地西他滨对白血病Kasumi-1细胞的毒性作用及其机制研究

Cytotoxicity and its mechanism of decitabine on Kasumi-1 cells in leukemia

目的:探讨地西他滨对白血病Kasumi-1细胞的毒性作用及其机制。方法:应用CCK-8法检测0.5～100μmol/L地西他滨作用Kasumi-1细胞24、48、72 h后的细胞存活率,An-nexinⅤ-FITC/PI双染色法检测0.5～20μmol/L地西他滨处理Kasumi-1细胞48 h后的细胞凋亡率,流式细胞碘化丙啶单染色法观察G0/G1期、S期、G2/M期Kasumi-1细胞比例;逆转录PCR法检测p21WAF1/CIP1基因表达水平的变化。结果:0.5～100μmol/L地西他滨均可抑制Kasumi-1细胞增殖,并呈现剂量依赖性与时间依赖性,作用24、48、72 h的IC50浓度分别为6.92、5.03、4.47μmol/L;0.5～20μmol/L地西他滨诱导Kasumi-1细胞凋亡水平较低,但能以剂量依赖的方式阻滞Kasumi-1细胞G2/M期,上调p21WAF1/CIP1基因表达。结论:地西他滨对Kasumi-1细胞具有毒性作用,其机制可能与上调p21WAF1/CIP1表达以及阻滞细胞周期有关。

Objective： To investigate the cytotoxicity and its mechanism of decitabine （DAC） on Kasumi＇l cells in leukemia. Methods： Cell viability rates of Kasumi-1 cells were determined by CCK-8 assay after treatment of DAC （0.5 - 100 μmol/L） for 24, 48 and 72 h. Apoptosis analysis was conducted by Annexin V/PI apoptotic detection assay on Kasumi-1 cells treated with 0. 5 - 20μmol/L DAC for 48 h. Cell cycle changes were assessed by flow cytometry （PI staining）. Reverse transcription-PCR （RT-PCR） was used to determine the mRNA expression levels of p21^WAF1/CIP1 gene. Results： DAC （0. 5 -100 μmol/L） significantly inhibited the growth of Kasumi-1 cells in both time- and dose- dependent manners. The ICs0 of DAC on the viability of Kasumi-1 cells 24, 48 and 72 h after treatment were 6. 92, 5.03 and 4.47 μmol/L, respectively. DAC （0. 5 -20 μmol/L） induced Kasumi-1 cell apoptosis, and further arrested G2/M cell cycle and up-regulated p21^WAF1/CIP1 gene in a dose-dependent manners. Conclusion： DAC exerts its cytotoxic effects on Kasumi-1 cells, accompanied with G2/M cell cycle arrest and up-regulation of p21^WAF1/CIP1.