人脐带间充质干细胞对依托泊苷诱导的Nalm-6细胞凋亡的影响

Effect of Human Umbilical Cord Mesenchymal Stem Cells on Etoposide-induced Nalm-6 Cell Apoptosis

目的:探讨人脐带间充质干细胞(hUC-MSC)对依托泊苷(VP16)诱导的B淋巴细胞白血病细胞系Nalm-6凋亡的影响。方法:首先分离出hUC-MSC,对其形态和免疫表型进行鉴定。然后利用VP16诱导Nalm-6细胞部分凋亡,将Nalm-6与hUC-MSC共培养,应用流式细胞术检测Nalm-6细胞周期及其凋亡变化,定量RT-PCR方法检测Nalm-6细胞中凋亡相关基因BCL-2、BAX和caspase-3的mRNA的表达变化,Western blot方法检测BCL2、BAX和caspase-3蛋白的表达变化。结果:流式细胞术检测显示,hUC-MSC能够抑制Nalm-6细胞增殖,减少VP16导致的Nalm-6细胞凋亡比例;RT-PCR检测表明,在VP16存在时,hUC-MSC增加Nalm-6细胞中BCL-2 mRNA的表达,降低BAX和caspase-3 mRNA的表达,差异均具有统计学意义(P<0.05);Western blot检测表明,hUC-MSC增加Nalm-6细胞中BCL-2蛋白的表达,降低BAX和caspase-3蛋白的表达,差异均具有统计学意义(P<0.05)。结论:hUCMSC可能通过上调BCL-2、下调BAX和caspase-3表达减少VP16诱导的Nalm-6细胞凋亡,从而对Nalm-6细胞产生保护作用。

Objective:To investigate the effect of human umbilical cord mesenchymal stem cells(hUC-MSC) on VP16-induced apoptosis of Nalm-6 cells.Methods:hUC-MSC were isolated and identified using morphological observation and flow cytometry,then Nalm-6 cells were treated with hUC-MSC with or without VP16.Apoptosis and cell cycle were assayed by FACS.The mRNA levels of apoptosis-related genes BCL-2,BAX and caspase-3 were detected by quantitative RT-PCR,and the protein levels of BCL-2,BAX and caspase-3 were examined by Western blot.Results:FACS showed that hUC-MSC inhibited the proliferation and decreased apoptosis of Nalm-6 cells resulted from VP16.The quantitative RT-PCR showed that hUC-MSC increased the mRNA expression level of BCL-2 and decreased the expression level of BAX and caspase-3(P <0.05).Western blot showed that the protein expression level of BCL-2 increased,and expression level of BAX and caspase-3 decreased in Nalm-6 cells after co-culture with hUC-MSC(P <0.05).Conclusion:hUC-MSC may protect Nalm-6 cells from apoptosis induced by VP16 through regulation of BCL-2,BAX and caspase-3.