糖基化磷脂酰肌醇特异性磷脂酶D对髓系白血病骨髓细胞黏附功能的影响及其机制(英文)

Effect and mechanism of glycosylphosphatidilinoditol-specific phospholipase D on the adhesion function of bone marrow mononuclear cells separated from myeloid leukemia patients

背景:据作者查新检索medline,cnki等数据库,鲜见国内外有关体外研究糖基化磷脂酰肌醇特异性磷脂酶D(glycosylphosphatidilinoditol-specific phospholipaseD,GPI-PLD)对白血病细胞黏附功能影响方面的报道。目的:观察GPI-PLD对髓系白血病骨髓单个核细胞黏附功能的影响及其机制。设计、时间及地点:细胞学体外实验,于2004-01/06在湘雅医院血液实验室完成。对象:骨髓标本来自于髓系白血病患者,由湘雅医院血液科提供。方法:利用具有完整糖基化磷脂酰肌醇(GPI)结构的胎盘碱性磷酸酶做底物,通过TX-114分相,定量检测骨髓单个核细胞中GPI-PLD活性,利用1,10-二氮杂菲抑制GPI-PLD的活性,实验分2组:处理组加二氮杂菲,使其终浓度为1mmol/L,对照组加入等量的PBS。用MTT方法检测处理组和对照组骨髓细胞对纤维连接蛋白的黏附率、免疫组织化学方法检测GPI-锚定蛋白CD24的表达。主要观察指标:髓系白血病细胞GPI-PLD活性,细胞黏附率及CD24的表达。结果:1mmol/L1,10-二氮杂菲作用髓系白血病骨髓单个核细胞5h细胞的GPI-PLD活性较对照组降低[(5.40±2.96)%,(42.08±7.21)%,P<0.01];GPI-PLD的活性被抑制后处理组细胞的黏附率较对照组增加[(61.19±29.14)%,(49.78±26.73)%,P<0.01],同时CD24的表达率上调[(18.5±11.14)%,(16.02±9.68),P<0.01]。结论:降低GPI-PLD活性能增加骨髓单个核细胞对纤维连接蛋白的黏附率,同时骨髓单个核细胞上GPI-锚定蛋白CD24表达增强。

BACKGROUND： There still is rarely report about the effect of glycosylphosphatidilinoditol-specific phospholipase D （GPI-PLD） on the adhesion function of leukemic cells through screening Medline and CNKI databases. OBJECTIVE： To observe the effect of GPI-PLD on the adhesion function of bone marrow mononuclear cells separated from myeloid leukemia patients, and to investigate the related mechanism. DESIGN, TIME AND SETTING： This study addressing cytology in vitro was conducted at the Hematological Laboratory of Xiangya Hospital from January to June 2004. MATERIALS： Bone marrow was collected from myeloid leukemia patients at the Department of Hematology, Xiangya Hospital, China. METHODS： The GPI-PLD activity of bone marrow mononuclear cells separated from myeloid leukemia patients was measured by using GPI-anchored placental alkaline phosphatase as substrate and Triton-X114 partition. By use of 1,10-phenanthroline, the activity of GPI-PLD was inhibited, the experiment was divided into 2 groups： treatment group adding phenanthroline to achieve a final concentration of 1 mmol/L, while control group adding the same amount of phosphate buffered saline. The adhesion rate to the fibronectin and CD24 expression of these cells were measured by MTT and immunohistochemical method, respectively. MAIN OUTCOME MEASURES： GPI-PLD activity of myeloid leukemic cells, cell adhesion rate, CD24 expression were all measured. RESULTS： The GPI-PLD activity of bone marrow mononuclear cells separated from myeloid leukemia patients was inhibited significantly after these cells were treated by 1 mmol/L 1,10-phenanthroline for 5 hours compared with control groups [（5.40±2.96）%, （42.08±7.21）%, P 0.01]. At the same time, the adhesion rate of these cells were increased after the GPI-PLD activity was inhibited [（61.19±29.14）%, （49.78±26.73）%, P 0.01], and the CD24 expression was also up-regulated [（18.5±11.14）%, （16.02±9.68）, P 0.01]. CONCLUSION： The adhesion rate of bone marrow mononuclear cells separated from myeloid leukemia patients can be promoted by inhibiting GPI-PLD activity. At the same time, the CD24 expression of GPI-anchored proteins on bone marrow mononuclear cells is improved.