糖基化磷脂酰肌醇特异性磷脂酶D(GPI-PLD)对髓性白血病骨髓细胞黏附功能的影响及机制探讨

Effect of GPI-PLD on Adhesion Function of Bone Marrow Mononuclear Cell from Patients with Myeloid Leukemia and Its Mechanism

为了探讨糖基化磷脂酰肌醇特异性磷脂酶D(GPIPLD)对髓性白血病骨髓单个核细胞黏附功能的影响及其机制,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(PLAP)做底物,通过TX114分相,定量检测骨髓单个核细胞中GPIPLD活性,用1,10二氮杂菲抑制GPIPLD的活性,用MTT方法检测抑制组和非抑制组骨髓细胞对纤连蛋白的黏附率,用免疫组织化学方法检测GPI锚定蛋白CD24的表达。结果显示:1,10二氮杂菲(1mmol/L)作用骨髓单个核细胞5小时可使细胞的GPIPLD的活性由(42.08±7.21)%降为(5.4±2.96)%,GPIPLD活性被抑制后细胞的黏附率增加,由(49.78±26.73)%升为(61.19±29.14)%,与此同时,CD24的表达率上调,由(16.02±9.68)%升为(18.5±11.14)%。结论:降低GPIPLD活性能增加骨髓单个核细胞对纤连蛋白的黏附率,与此同时,骨髓单个核细胞上GPI锚定蛋白CD24的表达增强。

To explore the effect of glycosyl-phosphatidyl inositol-specific phospholipase D (GPI-PLD) on the adhesion function of bone marrow mononuclear cell from patients with myeloid leukemia and analyze its mechanism, the activity of GPI-PLD in bone marrow mononuclear cell from the patients were measured by using GPI-anchored placental alkaline phosphatase (PLAP) as substrate and Triton-X114 partitioning; the adhesion rate and CD24 expression of these cells were measured by MTT and immunohistochemical method respectively, when these cells were or were not treated by 1 mmol/L 1,10-phenanthroline for 5 hours. The results showed that the GPI-PLD activity of bone marrow mononuclear cells from the patients was significantly inhibited after being treated by 1 mmol/L 1,10-phenanthroline for 5 hours [(42.08±7.21)% vs (5.4±2.96)%], while the adhesion rate and the expression of CD24 of these cells were increased [(49.78±26.73)% vs (61.19±29.14)%, (16.02±9.68)% vs (18.5±11.14)%, respectively )]. It is concluded that depression of GPI-PLD activity can increase the adhesion rate of bone marrow mononuclear cells from the patients while the CD24 expression is enhanced.