地西他滨联合全反式维甲酸对SKM-1 MDS细胞株的体外研究

目的:探讨去甲基化制剂地西他滨和全反式维甲酸对MDS-RAEB细胞株SKM-1的体外影响及其机制。方法:SKM-1细胞经药物处理后,用台酚蓝拒染法观察SKM-1细胞生长曲线的变化;用硝基四氮唑蓝还原试验和流式细胞术观察药物对SKM-1细胞的诱导分化作用;用Annexin Ⅴ-FITC标记了解细胞的早期凋亡;用RT-PCR研究细胞DNMT3A和P15INK4B基因表达的变化。结果:地西他滨和ATRA抑制SKM-1细胞生长,增强SKM-1细胞的还原能力,增加细胞表面CD14、CD11b表达,促进细胞凋亡,两药联合应用具有协同作用;地西他滨可使SKM-1细胞P15INK4B mRNA表达增加,DNMT3A mRNA表达减少。结论:地西他滨和ATRA均可以促进SKM-1细胞凋亡和分化,二者有协同作用;地西他滨的作用可能与DNMT3A和P15INK4B基因表达有关。

The effects and the mechanisms of demethylating agent Decitabine and/or ATRA on SKM-1 cells in vitro

Objective:To explore the effects and the mechanisms of demethylating agent Decitabine and/or ATRA on SKM-1 cells in vitro.Method:The proliferation of SKM-1cells was assessed by trypan blue exclusion staining;the differentiation of SKM-1cells was tested by nitro-blue tetrazolium(NBT) reduction and flow cytometry;the early apoptosis of the cells was detected by Annexin Ⅴ-FITC double staining;the mRNA expressions of DNMT3A and P15INK4B in the cells were assayed by reverse transcription-polymerase chain reaction(RT-PCR).Result:Decitabine and ATRA inhibited SKM-1cells' growth enhanced the cells' reducing power,increased the expressions of CD14 and CD11b,and induced apoptosis of the cells,Combination of Decitabine and ATRA had synergistic effect.Decitabine could up-regulate P15INK4B mRNA expression,and down-regulate DNMT3A mRNA expression.Conclusion:Both Decitabine and ATRA can induce the apoptosis and differentiation of SKM-1 cells,and their combination has synergistic effect.The effects of Decitabine may be linked to the expressions of DNMT3A and P15INK4B.