磷酸氟达拉滨对MUTZ-1细胞作用及机制研究

Inhibition effect of fludarabine on the growth of myelodysplastic syndrome cell line MUTZ-1 and its mechanism

目的:在体外研究磷酸氟达拉滨(fludarab ine)对人骨髓增生异常综合征细胞株MUTZ-1细胞的作用以及可能的作用机制。方法:采用MTT法、透射电境、DNA凝胶电泳、流式细胞仪以及RT-PCR等分析方法。结果:磷酸氟达拉滨能抑制MUTZ-1细胞的生长,24、48、72 h的IC50分别为137.65 m g/L、6.27 m g/L、0.51 m g/L,具有浓度和时间依赖性。经透射电境、DNA凝胶电泳和A nnex inⅤ/P I检测,证实1~16 m g/L磷酸氟达拉滨对MUTZ-1细胞作用24 h后,可以诱导MUTZ-1细胞凋亡,其对Bcl-2、Bax、surv iv in、X IAP、cIAP1和cIAP2基因mRNA表达均无明显下调作用,但可以导致MUTZ-1细胞线粒体膜电位的下降。结论:1 m g/L^16 m g/L磷酸氟达拉滨不但能抑制MUTZ-1细胞生长而且能诱导细胞凋亡,诱导细胞凋亡是其主要细胞毒作用之一。线粒体膜电位下降是其诱导细胞凋亡的重要环节之一。

Objective： To investigate the inhibition effect of fludarabine on the growth of human MDS-RAEB cell line MUTZ-1 and to explore the possible cellular and molecular mechanism. Methods： The apoptosis of MUTZ-1 cells induced by fludarabine was studied by transmission electron microscope, MTT assay, DNA ladder test, flow cytometry and RT-PCR method. Results： Treatment with fludarabine remarkably inhibited the growth of MUTZ-1 cells, the 24 h IC50, 48 h IC50 and 72 h IC50 of fludarabine for MUTZ-1 cells were 137.65 mg/L, 6.27 mg/L and 0.51 mg/L, respectively. Fludarabine inhibited the growth of MUTZ-1 cells in a dose-dependent and time-dependent manner. After treated by fludarabine （1 mg/L-16 mg/L） for 24 h, MUTZ-1 cells showed the typical features of apoptosis. After fludarabine treatment the mRNA expression of Bcl-2, Bax, survivin, XIAP, cIAP-1 and cIAP-2 was not changed, but the mitochondrial membrane potential （MMP） was decreased. Conclusion： With a certain range of dose fludarabine （1 mg/L-16 mg/L）could inhibit MUTZ-1 cell growth by inducing cells apoptosis. MMP may play a certain role in apoptosis of MUTZ-1 cells induced by fludarabine.