两条IL-6信号转导途径在人骨髓瘤细胞系U266中的相互拮抗作用

Antagonistic effect between two IL-6 signal transduction pathways in a human myeloma cell line-U266

目的 研究人骨髓瘤细胞系U2 6 6中的白细胞介素 6 (IL 6 )信号转导途径及彼此之间的相互调控方式。方法 首先采用凝胶阻滞电泳 (electrophoreticmobilityshiftassay,EMSA)方法观察分别参与两条IL 6信号转导途径的转录因子STAT3和NF IL 6在U2 6 6细胞中的诱导激活状态并确定该细胞中的IL 6信号转导通路 ;继而采用转基因或化学试剂处理方法 ,特异性上调或下调其中一条IL 6信号途径的活化 ,并同时观察另外一条信号途径的激活状态。结果 1.以上两种转录因子分别参与的IL 6信号转导途径———JAK/STAT和Ras/NF IL 6 ,它们都能够在U2 6 6细胞中诱导激活 ;2 .在高剂量IL 6 (1～ 10 0ng/ml)刺激范围之内 ,一条IL 6信号途径活化水平的升高可同时导致另一条信号途径活化水平的下降。结论 在一定IL 6刺激剂量范围内 ,U2 6 6细胞中两条IL 6信号途径的诱导激活存在着相互拮抗作用。

Objective To investigate the IL-6 signal transduction pathways and their regulation mechanism in a human myeloma cell line-U266. Methods Electrophoretic mobility shift assay (EMSA) was used to detect the activation of the transcription factors (TFs)-STAT3 and NF-IL-6 by IL-6. Cells were treated with chemical agents or transfected with the expression plasmids for the two TFs or the anti-sense oligonucleotide for protein kinase involved in one IL-6 signal transduction pathway. The change of the activation state of another IL-6 signal transduction pathway was also exhibited by EMSA. Results Two IL-6 signal transduction pathways (JAK/STAT and Ras/NF-IL-6) can be activated by IL-6 of different dose in U266 cells. When one of the two signal transduction pathways was up-regulated, the other one was down-regulated. Conclusion There is an antagonistic effect between the activation of two IL-6 signal transduction pathways in U266 cells.