摘要
目的观察急性T淋巴细胞白血病(T-ALL)患者骨髓CD+34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CD+34细胞比例及其细胞周期,免疫磁珠法分选CD+34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CD+34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CD+34细胞Hes1基因表达分别为3.3±0.8、1,两者比较,P<0.05;CD+34细胞比例分别为0.02%±0.003%、0.06%±0.005%,两者比较,P<0.05;CD+34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CD+34细胞的体外集落形成减少。提高正常CD+34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CD+34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 CD34^＋ cells were analyzed by FACS, CD3＋4 cells were cultured in vitro for colony formation cells （ CFC） .After increasing the expression of Hes1 in CD34^＋ 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 CD34^＋ 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 CD34^＋ 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.
出处
《山东医药》
CAS
2014年第39期5-7,共3页
Shandong Medical Journal
基金
国家自然科学基金项目(31301161)
天津市应用基础与前沿技术研究计划(13JCYBJC22800)
