摘要目的研究蛋白激酶ERK对人骨髓瘤细胞系Sko-007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-007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-007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-007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-007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.

abstractsObjective　To investigate the regulation effect of protein kinase ERK on the activation of transcription factor STAT3 in response to IL-6 in the Sko-007 human myeloma cell line.Methods　Electrophoretic mobility shift assay (EMSA) and immunoprecipitation (IP) were used to show the activation of STAT3 and ERK in Sko-007 cells in the presence and absence of IL-6. Antisense oligonucloetides of ERK (ERK-AS) were transfected into Sko-007 cells to specifically inhibit the expression and activity of ERK. The changes in the activation of STAT3 in the transfected cells were also exhibited by EMSA. Direct binding between STAT3 and ERK was analyzed by co-IP.Results　Both STAT3 and ERK were activated in Sko-007 cells stimulated with IL-6. ERK-AS inhibited STAT3 activation by IL-6. Moreover, activated ERK could form a complex with STAT3 in Sko-007 cells.Conclusion　ERK can bind STAT3 directly and be required for its maximal activation in Sko-007 cells stimulated by IL-6.