摘要
目的 探讨白细胞介素 6 (IL 6 )基因在不同类型白血病细胞中的构成表达以及白血病细胞是否自泌IL 6 ,为更好地利用IL 6 /IL 6R系统介导重组白细胞介素 6 假单胞菌外毒素融合蛋白(IL 6 PE40 )靶向治疗白血病提供可靠依据。方法 采用反转录 聚合酶链反应 (RT PCR)半定量技术、序列分析及ELISA方法检测白血病细胞系U937、HL6 0、KG1、TF1、K5 6 2、HuT2 8、CEM及Raji中IL 6mRNA的表达以及IL 6的自泌含量。结果 IL 6mRNA在粒、红、单以及淋系白血病细胞系中的相对表达丰度甚低 ,仅在 0 0 3～ 0 0 7之间 ,而慢性粒细胞白血病K5 6 2却高表达 ,其丰度达 1 2 5 ;所检测的8种白血病细胞在自泌IL 6水平上也存在着显著差异 ,按分泌含量的不同拟可将各型白血病细胞分为高、中、低三型。结论 不同类型的白血病细胞其IL 6基因的构成性表达和自泌IL 6的水平是明显不同的 ,大量自泌的IL 6有可能会对重组IL 6 PE40的靶向性结合产生竞争性抑制 ,从而影响其特异性杀伤白血病细胞的效果。
Objective To make a comprehensive and systematic study on the constructive expression of IL 6 mRNA and IL 6 autosecretion in human leukemic cells and discuss the effect of IL 6 autosecretion on the specific binding of IL 6 PE40 fusion protein to targeted leukemic cells. Methods Semi quantitative RT PCR,sequencing and ELISA were used to detect the constructive expression of IL 6 mRNA and autosecretion level of IL 6 protein in human leukemic cell lines, such as U937、HL60、KG1、TF1、K562、HuT28、CEM and Raji. Results Therelative expression level of IL 6 mRNA in myelocytic,monocytic and erythrocytic leukemic cell lines including HL60,U937,KG1 and TF1 and lymphoblastic leukemic cell lines such as CEM,HuT28 and Raji is very weak,ranging from 0.03 to 0.07. However, K562, an chronic myeloecytic leukemic cell line, had the highest level 1.25 among the 8 leukemic cell lines and the level was also higher than that of the positive control U266 multiple myeloma cell line. Based on ELISA assay of IL 6, there were remarkable differences between the 8 leukemic cell lines, their secretion of IL 6 could be divided into 3 levels ,i.e. high, moderate and low secretion type. Conclusion These observations imply that the intrinsic expression and secretion of IL 6 differ obviously among different leukemic cell lines. Autocrine IL 6 may prevent IL 6 PE40 fusion protein from specific binding to targeted leukemic cells and consequently affect its specific killing toxicity.
出处
《中华内科杂志》
CAS
CSCD
北大核心
2000年第9期611-613,I021,共4页
Chinese Journal of Internal Medicine
基金
国家自然科学基金!资助项目 (395 0 0 0 6 2和 39870 875 )
