gp130胞外区近膜端缺失的突变体介导IL-6信号的研究

目的研究ｇｐ１３０胞外区远膜端和近膜端在ＩＬ－６信号转导中的作用。方法用分子克隆手段构建全长的膜结合型ｇｐ１３０分子和胞外区第３０１～５８２位氨基酸残基缺失的突变体分子，并通过阻滞电泳方法比较突变体与野生型分子在传导信号方面的差异。结果通过酶切鉴定、序列分析和表达检测，证明野生型和突变体ｃＤＮＡ表达载体构建正确，并有效表达；阻滞电泳结果表明，在ＳＫＯ－００７细胞和Ｍｏｌｔ－４细胞中，突变体分子与野生型ｇｐ１３０都能介导ＩＬ－６信号。结论ｇｐ１３０分子胞外区远膜端在ＩＬ－６信号转导中发挥重要作用；而近膜端并非ＩＬ－６信号转导所必需。

Studies on IL 6 signals transduced by gp130 deleted proximal membrane part of extracellular region

Objective To study the functions of the distal membrane part and the proximal membrane part of gp130 extracellular region in IL 6 signal transduction. Methods A wild type membrane anchored gp130 and a membrane anchored gp130 mutant,without amino acids 301 582 in the proximal membrane part of gp130 extracellular region were constructed with molecular biological techniques. The function differences of the two molecules in IL 6 signal transduction were compared with electrophoresis mobility shift assays (EMSA). Results The two gp130 cDNA vectors constructed by us were manifested to be correct by restriction digestion and sequencing,and to express their specific mRNA in target cell lines demonstrated by dot blot method.The function of gp130 mutant was detected to be similar to that of wild type gp130 in transducing IL 6 signals in SKO 007 cell line and Molt 4 cell line by EMSA. Conclusion These results suggest that the distal membrane part of gp130 extracellular region is crucial in IL 6 signalling,while the proximal membrane part is not necessary.