胚胎干细胞对急性髓系白血病细胞KG-1a的影响

Effect of Human Embryonic Stem Cells on Acute Myeloid Leukemia KG-1a Cells

目的:研究人胚胎干细胞H9在体外对人急性髓系白血病细胞KG-1a增殖、凋亡的影响,并探讨其作用机制。方法:体外将人胚胎干细胞H9和白血病细胞KG-1a进行接触式共培养,用CCK8检测KG-1a的增殖情况;用流式细胞术检测共培养后细胞的凋亡以及周期改变;荧光定量PCR检测BAX、BCL-2、caspase-3的m RNA表达;Western blot分析BAX、BCL-2、caspase-3的蛋白表达。结果:H9能明显抑制KG-1a细胞的增殖,将细胞阻滞于G2/M期,共培养后的KG-1a细胞的凋亡率高于对照组。BAX、Caspse-3的m RNA和蛋白表达均上调,BCL-2的m RNA和蛋白表达下调。结论:胚胎干细胞能抑制KG-1a细胞的增殖,促进其凋亡,其机制可能与BCL-2表达下调和BAX、Caspse-3表达上调有关。

Objective： To investigate the effect of embryonic stem cells on the proliferation and apoptosis in human acute myeloid leukemia cell line KG-1 a and to explore its potential mechanism. Methods： The direct co-culture system between human embryonic stem cells H9 and human acute myeloid leukemia cell line KG-la was established, and CCK8 assay was used to detect the proliferation of KG-1 a cells. The changes of cell cycle and apoptosis were detected by flow cytometry （FCM）. The mRNA expressions of BCL-2, BAX, Caspase-3 were assessed by RT-PCR. Meanwhile, the protein-expressions of BCL-2, BAX, Caspase-3 were detected by Western blot. Results ： The proliferation level of KG- 1a cells was significantly inhibited by Hg, and the apoptotic rate increased, and the cell cycle was blocked at G2/M phase. The mRNA-expression and the protein-expression of BAX and Caspase-3 increased, the mRNA and protein- expression of BCL-2 decreased. Conclusion： Embryonic stem cells can inhibit the proliferation of KG-1 a and induce the apoptosis that maybe relate with the down-regulation of BCL-2 expression and up-regulation of BAX and caspase-3 expression.