Hes1对急性髓系白血病患者骨髓CD34^＋CD38^-细胞的作用及其机制研究

Effect of Hes1 on bone marrow CD34^+ cells in acute myeloid leukemia

目的:研究Hes1对急性髓系白血病(AML)患者骨髓CD34+CD38-细胞的作用及其机制。方法:收集初治AML患者及正常供者骨髓样本后,通过密度梯度离心法获取单个核细胞,流式细胞术检测CD34+CD38-细胞比例及其细胞周期。通过免疫磁珠法分选CD34+CD38-细胞后,体外集落形成实验(CFC)检测其增殖能力,并通过Realtime PCR检测其Hes1的表达量。构建Hes1过表达逆转录病毒载体,感染正常供者骨髓CD34+细胞后,流式细胞术分析其细胞周期的改变,CFC检测其增殖的改变。结果:AML患者骨髓CD34+CD38-细胞比例明显低于正常对照,流式细胞术结果显示患者来源CD34+CD38-细胞大多数进入静止期,CFC结果显示患者CD34+CD38-细胞体外扩增能力下降。Realtime PCR结果发现患者CD34+CD38-细胞中Hes1表达上调。提高正常供者CD34+细胞中Hes1的表达后,细胞增殖减少,进入静止期。结论:在AML中CD34+CD38-细胞比例下降,进入静止期,与Hes1的表达上调有关。

Objective： To determine the effect of Hesl on bone marrow CD34＋ cells in acute myeloid leukemia （AML）. Meth- ods： Bone marrow mononuclear cells were isolated by using Ficoll. Then, the proportion and cell cycle of CD34＋ cells were analyzed by using fluorescence-activated cell sorting （FACS）. CD34~ cells were cultured in vitro for colony-forming cells （CFC）. The expression of Hes 1 in CD34＋ cells was evaluated by using real-time polymerase chain reaction. After upregulating the expression of Hes 1 in CD34＋ cells, the cell cycle was analyzed through FACS, and the colony formation of CD34~Hes 1 ＋ cells was analyzed by CFC. Results： The ra- tio of CD34~ cells in the bone marrow was lower in the AML group than in the control group. In addition, more CD34~ cells underwent quiescence in the AML group than in the control group. In vitro assay showed that the colony formation of CD34＋ cells was lower in the AML group than in the control group. The expression of Hesl was higher in the CD34＋ cells from the AML patients than that in the CD34＋ ceils from normal donors. After Hesl transduction, more CD34＋ ceils underwent quiescence and showed weak proliferation, Conclusion： The proportion of CD34＋ cells in the bone marrow was lower in AML patients than in normal donors. A large proportion of CD34＋ cells underwent quiescence, which was related to Hes 1, in AML patients.