T-ALL 患者骨髓 CD3＋4细胞的 Hes1基因表达、数量、增殖变化及机制探讨

摘要

目的：观察急性T淋巴细胞白血病（ T-ALL）患者骨髓CD3＋4细胞Hes1基因表达、数量、增殖的变化，并探讨其机制。方法收集8例初治T-ALL患者及4例正常供者（健康对照）骨髓样本，real time PCR检测Hes1基因的表达，密度梯度离心法获取单个核细胞，流式细胞术检测CD3＋4细胞比例及其细胞周期，免疫磁珠法分选CD3＋4细胞，体外集落形成实验（ CFC）检测其增殖能力。构建Hes1基因过表达逆转录病毒载体，感染正常供者骨髓CD3＋4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 T-ALL患者、健康对照CD3＋4细胞Hes1基因表达分别为3．3±0．8、1，两者比较，P＜0．05；CD3＋4细胞比例分别为0．02％±0．003％、0．06％±0．005％，两者比较，P＜0．05；CD3＋4细胞处于S期的细胞比例分别为16．2％±0．98％、28．0％±1．12％，两者比较，P＜0．05；G0期比例分别为19．0％±0．9％、9．0％±0．5％，两者比较，P＜0．05；且患者来源CD3＋4细胞的体外集落形成减少。提高正常CD3＋4细胞中Hes1基因的表达后，细胞增殖减少，进入静止期。结论 T-ALL患者CD3＋4细胞比例下降，进入静止期，体外扩增能力下降，这可能与Hes1基因的表达上调有关。%Objective To study the expression of Hes1, cell cycle, cell proliferation of bone marrow CD3+4 cells in acute T lymphoblastic leukemia( T-ALL) patients.Methods Samples of 8 T-ALL patients and 4 healthy donars were collected. Hes1 expression was analyzed by real time PCR.Bone marrow mononuclear cells were isolated with Ficoll and then the pro-portion and cell cycle of CD3+4 cells were analyzed by FACS, CD3+4 cells were cultured in vitro for colony formation cells ( CFC) .After increasing the expression of Hes1 in CD3+4 cells, cell cycle was analyzed by FACS and the colony formation of CD3+4 cells were analyzed by CFC.Results Hes1 expression in T-ALL and control were 3.3 ±0.8 and 1 respectively(P<0.05).The ratio of CD3+4 cells in T-ALL and control were 0.02%±0.003%and 0.06%±0.005%respectively(P<0.05). The proportion of S phase of CD3+4 cells in T-ALL and control were 16.2%±0.98%and 28.0%±1.12%respectively(P<0.05).The proportion of G0 phase of CD3+4 cells were 19.0%±0.9%and 9.0%±0.5%respectively(P<0.05).The colo-ny formation of CD3+4 cells from patients was lower than that from normal donors.More CD3+4 cells went into quiescence and the proliferation of these cells were inhibited after overexpressing Hes1.Conclusions In T-ALL patients, the proportion of CD3+4 cells in bone marrow is lower.More of these cells go into quiescence which is related to Hes1.