慢病毒介导的GFI-1基因沉默对慢性髓细胞白血病患者单个核细胞生长及凋亡的影响  

Impact of lentivirus- mediated gene silencing of growth factor independence- 1 in chronic myeloid leukemia mononuclear cells

作　　者：杨熙[1] 刘红[1] 林赠华[1] 陆伟[1] 张亚平[1] 

机构地区：[1]南通大学附属医院血液科,江苏南通226001

出　　处：《广东医学》2016年第13期1944-1947,共4页Guangdong Medical Journal

基　　金：国家自然科学基金资助项目(编号:81070400);江苏省医学创新团队与领军人才项目(编号:LJ201136);江苏省南通市社会事业科技创新与示范计划项目(编号:HS2013067)

摘　　要：目的观察慢病毒介导的GFI-1基因沉默后对慢性髓细胞白血病患者单个核细胞生长及凋亡的影响,研究GFI-1在慢性髓细胞白血病发病中作用。方法收集16例慢性髓细胞白血病初治患者的外周血标本,以17例正常人的外周血作为正常对照。并采集患者及正常人的临床相关资料,以慢病毒感染方法沉默GFI-1基因,以实时定量PCR、Western blot、流式细胞术等方法研究慢性髓细胞白血病患者GFI-1在单个核细胞中的表达及基因沉默后对细胞生长及凋亡的影响。结果 GFI-1在慢性髓细胞白血病初治患者中的表达明显高于正常人,差异有统计学意义(P<0.001)。GFI-1沉默后72 h,与感染了空载体的慢性髓细胞白血病单个核细胞相比细胞增殖显著受抑制(P<0.001),细胞凋亡也明显高于感染了空载体的慢性髓细胞白血病单个核细胞(P<0.005),细胞周期阻滞。结论 GFI-1可能通过调控凋亡通路及细胞周期相关因子导致细胞凋亡周期阻滞,从而抑制细胞增殖。Objective To investigate effects of lentivirus - mediated gene silencing of growth factor independence - 1 （GFI- 1 ） on the growth and apoptosis of mononuclear cells in chronic myelogenous leukemia（CML） patients, thus to e- valuate the significance of GFI - 1 in pathogenesis of CML. Methods Peripheral blood and bone marrow samples were collected from 16 CML patients. As control group, another 17 samples were collected from normal healthy people. The ex- pression of GFI - 1 in mononuclear cells in CML patients was silenced by using shRNA. After that the proliferative ability of mononuclear cells were tested by MTS assay, as apoptosis was examined by AnnexinV -propidium iodide（PI） staining, and DNA content was measuredby PI staining. Results GFI - 1 expression was up - regulated in CML patients and significantly higher than that in control group （P 〈 0. 001 ）. GFI - 1 gene silencing can significantly inhibit mononuelear cells proliferation in 72 h （P 〈0. 001 ） and induce apoptosis in mononuclear cells （P 〈 0. 005 ）, which caused the retardation of cell cycle in mononuclear cells. Conclusion GFI - 1 may contribute to inhibit cell proliferation by leading to apoptosis and retardation of cell cycle as a proliferation regulator.

关 键 词：慢性髓细胞白血病 GFI-1 单个核细胞 慢病毒转染 细胞增殖 

分 类 号：R733.72[医药卫生—肿瘤]