CAJ | 학술논문

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。
목적：연구Hes1대급성수계백혈병（AML）환자골수CD34+CD38-세포적작용급기궤제。방법：수집초치AML환자급정상공자골수양본후，통과밀도제도리심법획취단개핵세포，류식세포술검측CD34+CD38-세포비례급기세포주기。통과면역자주법분선CD34+CD38-세포후，체외집락형성실험（CFC）검측기증식능력，병통과Realtime PCR검측기Hes1적표체량。구건Hes1과표체역전록병독재체，감염정상공자골수CD34+세포후，류식세포술분석기세포주기적개변，CFC검측기증식적개변。결과：AML환자골수CD34+CD38-세포비례명현저우정상대조，류식세포술결과현시환자래원CD34+CD38-세포대다수진입정지기，CFC결과현시환자CD34+CD38-세포체외확증능력하강。Realtime PCR결과발현환자CD34+CD38-세포중Hes1표체상조。제고정상공자CD34+세포중Hes1적표체후，세포증식감소，진입정지기。결론：재AML중CD34+CD38-세포비례하강，진입정지기，여Hes1적표체상조유관。
Objective:To determine the effect of Hes1 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 Hes1 in CD34+cells was evaluated by using real-time polymerase chain reaction. After upregulating the expression of Hes1 in CD34+cells, the cell cycle was analyzed through FACS, and the colony formation of CD34+Hes1+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 Hes1 was higher in the CD34+cells from the AML patients than that in the CD34+ cells from normal donors. After Hes1 transduction, more CD34+ cells 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 Hes1, in AML patients.