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目的 研究蛋白激酶ERK对人骨髓瘤细胞系Sko 0 0 7中转录因子STAT3在IL 6刺激下诱导活化的调控作用。方法 首先分别采用凝胶阻滞电泳 (EMSA)和免疫沉淀 (IP)方法观察IL 6刺激前后Sko 0 0 7细胞中STAT3和ERK的诱导活化情况。然后将ERK反义寡核苷酸 (ERK AS)转染入Sko 0 0 7细胞用来特异性抑制ERK的表达及功能 ,并同时观察STAT3诱导激活情况的改变。最后采用免疫共沉淀方法检测STAT3与ERK之间是否存在直接相互结合作用。结果 STAT3和ERK在Sko 0 0 7细胞中都能够被IL 6诱导激活 ;ERK AS转染后STAT3的诱导活化信号明显减弱。同时 ,STAT3和ERK可在IL 6刺激后发生直接相互结合作用。结论 Sko 0 0 7细胞中ERK可直接结合并参与IL 6刺激作用下STAT3的完全诱导活化。
Objective To study the regulation of protein kinase ERK on the activation of transcription factor STAT3 in human myeloma cell line Sko 0707 under IL-6 stimulation. Methods The activation of STAT3 and ERK in Sko 0 7 cells before and after IL 6 stimulation was observed by gel electrophoresis (EMSA) and immunoprecipitation (IP) methods, respectively. Then ERK antisense oligonucleotide (ERK AS) was transfected into Sko 0 07 cells to specifically inhibit the expression and function of ERK. At the same time, the activation of STAT3 was observed. Finally, the co-immunoprecipitation method was used to detect the direct interaction between STAT3 and ERK. Results Both STAT3 and ERK were activated by IL 6 in Sko 0 07 cells. The activation signal of STAT3 was significantly decreased after ERK AS transfection. At the same time, STAT3 and ERK can interact directly after IL 6 stimulation. Conclusion ERK can directly bind and participate in the complete induction and activation of STAT3 induced by IL-6 in Sko0.7 cells.