IL-6信号转导与转录激活子活性研究

Involvement of STAT3/APRF in IL 6 signal transduction pathway

目的 探讨 Ｉ Ｌ６ 生物效应与胞内磷酸化蛋白和转录激活子活性变化之间的关系。方法 应用原位杂交和 Ａ Ｐ Ａ Ａ Ｐ 方法检测细胞内 Ｉ Ｌ６ Ｒ ｍ Ｒ Ｎ Ａ 和蛋白的表达；采用 Ｄ Ｎ Ａ 结合蛋白凝胶阻滞电泳分析 Ｉ Ｌ６ 信号转导与 Ａ Ｐ Ｒ Ｆ 活性的相关性；用免疫沉淀和 Ｓ Ｄ Ｓ Ｐ Ａ Ｇ Ｅ 观察 Ｉ Ｌ６ 信号转导中磷酸化蛋白的变化情况。结果 （１） Ｉ Ｌ６ 受体（ Ｉ Ｌ６ Ｒ） ｍ Ｒ Ｎ Ａ 和蛋白表达阳性的人骨髓瘤细胞系（ Ｓ Ｋ Ｏ００７） 体外对 Ｉ Ｌ６ 有增殖反应；（２） 胞内转录激活子活性与 Ｉ Ｌ６ 诱导时间和剂量有相关性，抗ｇｐ１３０ 单克隆抗体和酪氨酸蛋白激酶抑制剂均可特异性抑制转录激活子活性；（３） Ｉ Ｌ６ 诱导 Ｓ Ｋ Ｏ００７细胞后，胞浆内出现一组相对分子质量为（１３０ 、９０ 、５４ 、３６） ×１０３ 磷酸化蛋白，它们的磷酸化活性与转录激活子活性一样，也具有时间和剂量相关性。结论 结果提示，转录激活子和这组相对分子质量不同的酪氨酸磷酸化蛋白参与 Ｉ Ｌ６ 信号转导调节。

Involvement of STAT3/APRF in IL6 signal transduction pathway$$$$ LI Yan, LAI Chunning, LIN Hong, et al. Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850 【 Abstract 】 Objective To explore the relationship between the biological effect of interleukin 6 (IL6) and intracellular phosphorylated proteins as well as the transducer and activator of transcription (STAT)/APRF. Methods In situ hybridization and APAAP were used to detect the expression of IL6R mRNA and protein in different cells, respectively. The correlation of IL6 signal transduction to the activity of APRF was determined by the DNA binding protein EMSA. The immunosedimentation and SDSPAGE were used to detect protein phosphorylation involved in IL6 signal transduction. Results IL6 stimulated the proliferation of human myeloma cell line (SKO007) expressing IL6R mRNA and proteins. Furthermore, IL6 showed a doseand timedependent activation of APRF and induced the phosphorylation of a group of intracellular proteins (130kD, 90kD, 54kD, and 36kD) in SKO007 cell line. The effect of IL6 on APRF activation could be blocked by an antigp130 monoclonal antibody and by the tyrosine kinase inhibitor, HA. Conclusion The results suggest that APRF and the tyrosinephosphorylated proteins play an important role in the regulation of IL6 signal transduction. 【 Subject words 】 Interleukin 6 Signal transduction Transcription activator Protein phosphorylation