基质细胞共培养对急性髓系白血病细胞耐药性影响及机制研究

Effect of Stromal Cell Co-culture on Drug Resistance of Acute Myeloid Leukemia Cells and its Mechanism

目的:研究基质细胞共培养对急性髓系白血病细胞耐药性的影响及机制。方法:将急性髓系白血病细胞HL-60与骨髓基质细胞HS-5共培养,用抗代谢药物阿糖胞苷(Ara-C)、蒽环霉素类药道诺霉素(DNR),生物碱类药物高三尖杉酯碱(HHT)对细胞进行联合处理,观察HL-60与HS-5共培养后对药物(DNR,HHT和Ara-C)敏感性的变化。同时用PI3K/AKT信号通路抑制剂LY294002处理细胞,研究共培养后细胞敏感性的变化与PI3K/AKT信号通路激活的相关性。结果:细胞抑制率的统计结果表明,HL-60与HS-5共培养后对药物的敏感性下降;mRNA定量和免疫蛋白印迹检测结果表明,HL-60与HS-5共培养后PI3K/AKT信号通路被激活,CCND1,FOXO1,PTEN等重要基因发生显著变化。结论:HL-60细胞通过与HS-5细胞共培养激活PI3K/AKT信号通路中的重要分子,如CCND1,FOXO1,PTEN,从而使HL-60细胞对药物的敏感性下降。

Objective： To study the effect of co-culture of stromal cells and acute myeloid leukemia（AML） cells on drug resistance of AML cells and its mechanism. Methods ： Stromal cells were co-cultured with acute myeloid leukemia cell HL-60 and then were treated with DNR, HHT and Ara-C for observing the sensitivity of HL-60 cells to drugs after incubation with HS-5. At the same time, the the inhibitor LY294002 of PI3K/AKT signaling pathway was used to treat the cells, so as to explore whether the changes of HL-60 sensitivity is associated with the activation of PI3K/AKT signal pathway after co-culture of cells. Results： The statistical results of HL-60 cell inhibition rate showed that the HL-60 cell sensitivity to drugs was decreased after incubation with HS-5, the mRNA quantitation and immunblot detection showed that PI3K/AKT signaling pathway was activated after co-culture of HL-60 cells with HS-5 cells, in addition the CCND1, FOXO1, PTEN and other important genes were also changed significantly. Conclusion： After co-culture of HL-60 cells with HS-5, some important molecules of PI3K/AKT signal pathway are changed, such as CCND1, FOXO1, PTEN, finally leading to the change of HL-60 cell sensitivity to drugs.