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目的 :研究各种白血病细胞膜IL 6R的表达、密度及亲和力 ,为深入探讨对IL 6 /IL 6R系统介导IL 6 PE40外毒素融合蛋白靶向杀伤白血病细胞的敏感性提供可靠依据。方法 :采用放射性配基结合实验 (RBA)并结合Scatchard作图法 ,系统分析IL 6R在 8种极具代表性的人类白血病细胞系U937,HL 6 0 ,KG1,TF1,K5 6 2 ,CEM ,HuT2 8和Raji上的表达密度及亲和力 ,同时应用流式细胞术 (FACS)检测IL 6R的α和 β亚单位蛋白在这些白血病细胞上的表达。结果 :RBA表明 ,粒、单、红白血病细胞系HL 6 0 ,U937,KG1和TF1表面存在着高亲和力IL 6R ,平均IL 6R密度 /细胞分别为 2 5 0 2个 ,2 874个 ,2 319个及 932 9个 ,而淋系白血病细胞系CEM ,HuT2 8和Raji以及慢粒K5 6 2细胞系则未测到IL 6R。FACS显示 ,HL 6 0 ,KG1,U937和TF1均高表达IL 6R的α亚单位蛋白 ,而CEM ,HuT2 8及K5 6 2则不表达IL 6Rα亚单位蛋白 ;IL 6Rβ亚单位蛋白在U937,KG1,TF1,HuT2 8及CEM中呈阳性表达 ,而在HL 6 0 ,Raji和K5 6 2细胞中则为阴性。结论 :鉴于粒、单、红白血病细胞异常高表达IL 6R ,而淋系和慢粒白血病呈阴性表达这一事实 ,提示急非淋白血病可能更适合用重组IL 6 PE40外毒素融合蛋白进行IL 6 /IL 6R介导的靶向治疗。
Objective: To study the expression, density and affinity of IL 6R in various leukemia cell membranes, and to provide a reliable basis for further investigation of the sensitivity of IL 6/IL 6R system-mediated IL 6 PE40 exotoxin fusion protein to killing leukemia cells. METHODS: Using radioactive ligand binding assay (RBA) combined with Scatchard mapping, the IL 6R was systematically analyzed in eight representative human leukemia cell lines U937, HL 60, KG1, TF1, K562, CEM, Expression densities and affinities on HuT28 and Raji, together with flow cytometric (FACS) detection of IL 6R alpha and beta subunit protein expression on these leukemia cells. RESULTS: RBA showed that there were high-affinity IL 6R on the surface of HL 60, U937, KG1, and TF1 cells in granulocyte, erythroleukemia and leukemia, and the mean IL 6R density/cells were 2,502, 2,874, and 2,319, respectively. And 9329, while the leukemia cell lines CEM, HuT28 and Raji, and the slow-grain K562 cell line did not detect IL6R. FACS showed that HL 6 0, KG1, U937 and TF1 all highly expressed IL 6R α subunit protein, while CEM, HuT28 and K562 did not express IL 6Rα subunit protein; IL 6Rβ subunit protein in U937, KG1 Positive expression was observed in TF1, HuT28, and CEM, but negative in HL60, Raji, and K562 cells. Conclusion: In view of the abnormally high expression of IL 6R in leukemic, erythroleukemia and leukemia cells, the fact that leukemia and chronic myeloid leukemia are negative indicates that acute non-leaf leukemia may be more suitable for IL 6 with recombinant IL 6 PE40 exotoxin fusion protein. /IL 6R-mediated targeted therapy.