Tetteroo P A, Massaro F, Mulder A, Schreuder-van Gelder R, von dem Borne A E
Leuk Res. 1984;8(2):197-206. doi: 10.1016/0145-2126(84)90143-7.
The human proerythroblastic leukemia cell-line K562 was induced to differentiate into megakaryocytic cells by 12-O-tetradecanoylphorbol-13-acetate (TPA). Megakaryocytic differentiation was detected when lineage-specific monoclonal antibodies were used to monitor the effect of TPA on K562 cells. A monoclonal anti-platelet antibody (C17) directed against an epitope present on GP IIIa appeared to react with K562 cells after induction. This was observed together with the disappearance of glycophorin A, the erythrocyte-specific lineage antigen. The induced megakaryocytic cells were also detected by ultrastructural platelet peroxidase (PPO). Immunoprecipitation, after ectolabeling of the cells with the C17 antibody and SDS-polyacrylamide gel electrophoresis, proved that TPA-induced K562 expressed both GP IIIa and GP IIb. However, the monoclonal antibody C15 directed against another epitope of platelet GP IIIa reacted only partially, or not at all, indicating that GP IIIa expressed on TPA-induced K562 differs structurally from that on normal platelets. K562 clones, expressing glycophorin A in all cells, were obtained by limiting dilution and culture. When these clones were treated with TPA, again megakaryocytic cells were obtained. These findings are discussed in relation to normal megakaryocytopoiesis.
人原红细胞白血病细胞系K562通过12 - O - 十四烷酰佛波醇 - 13 - 乙酸酯(TPA)诱导分化为巨核细胞。当使用谱系特异性单克隆抗体监测TPA对K562细胞的作用时,可检测到巨核细胞分化。一种针对糖蛋白IIIa(GP IIIa)上存在的表位的单克隆抗血小板抗体(C17)在诱导后似乎与K562细胞发生反应。这一现象与红细胞特异性谱系抗原血型糖蛋白A的消失同时被观察到。诱导的巨核细胞也通过超微结构血小板过氧化物酶(PPO)检测到。在用C17抗体对细胞进行细胞表面标记并进行SDS - 聚丙烯酰胺凝胶电泳后进行免疫沉淀,结果证明TPA诱导的K562细胞同时表达GP IIIa和GP IIb。然而,针对血小板GP IIIa另一个表位的单克隆抗体C15仅部分反应或根本不反应,这表明TPA诱导的K562细胞上表达的GP IIIa在结构上与正常血小板上的不同。通过有限稀释和培养获得了在所有细胞中都表达血型糖蛋白A的K562克隆。当用TPA处理这些克隆时,再次获得了巨核细胞。结合正常巨核细胞生成对这些发现进行了讨论。