粒单系白血病原始细胞表达血小板特异性抗原与周期蛋白D_3异常表达关系的研究

Abnormal Expression of Cyclin D_3 Causes Aberrant Expression of Platelet Specific Antigens on Myelomonocytic Leukemia Cells

目的 :探讨急性单核细胞白血病原始细胞表达血小板特异性抗原的机制。方法 :分离外周血或骨髓单个核细胞 ,采用常规染色和普通细胞化学染色观察其形态学特性 ,采用流式细胞仪双荧光分析免疫表型 ,单荧光胞浆蛋白标记分析周期蛋白表达和免疫印迹进一步证实周期蛋白 D3的表达情况。采用周期蛋白和 DNA双标记分析周期蛋白 D3表达与细胞周期的关系。结果 :白血病原始细胞中 CD1 3 + + HL A- DR+细胞为 94.6 9% ,CD1 3+ + CD34+细胞为 96 .86 % ,CD41 a+ + CD34+ 细胞为 41.6 0 % ,CD1 3 + + CD41 a+ 细胞为 40 .0 0 % ,免疫印迹和流式细胞仪单参数分析证实周期蛋白 D3表达明显升高 ,周期蛋白 D3阳性细胞在细胞周期各时段所占的比例分别为 :G1 期 5 2 .10 % ,S期9.90 % ,G2 + M期 38.0 0 %。结论 :周期蛋白 D3的异常表达导致单核系白血病细胞表达血小板特异性抗原 ,同时也可能在白血病的发生中起重要作用。

Objective: The aim of this investigation was to study the mechanism about the aberrant expression of platelet specific antigen on myelomyeloid leukemia cells. Methods: Mononuclear cells isolated from bone marrow were double labeled for membrane antigens, single labeled for cytoplasmic cyclins and, also stained with cyclin D 3 and propidium iodide for DNA. The treated cells were analyzed with FACSort flow cytometer. Immuno western were also performed to confirm the abnormal expression of cyclin D 3. Results: The percentage of CD 13 ++HLA DR +, CD 13 ++CD 34 +, CD 41a ++CD 34 +, and CD 41a ++CD 13 + cell are 94 69%, 96 86%, 41 60%, and 40 00%, respectively. Immuno western blot and flow cytometry showed the most of leukemia cells expressed cyclin D 3. Analysis of cell cycle distribution showed that percentage of G 1, S, and G 2+M phase of cyclin D 3 positive leukemia cells are 52.10%, 9.90, and 38.00%, respectively. Conclusion: The aberrant expression of platelet specific antigens on myelomonocytic leukemia cells is due to the abnormal expression of cyclin D 3. Additionally because the expression of cyclin D 3 increased significantly in G 2+M phase, the abnormal expression of cyclin D 3 could also play an important role in the development of leukemia.