摘要
目的 探讨只含白细胞介素 6 (IL 6 )受体 β亚基 (gp130 )胞外第二、三个FNⅢ (CBD)区的膜结合型gp130突变体 (mgp130 )能否传导IL 6信号。方法 首先用重叠延伸PCR方法扩增出mgp130cDNA ,将其克隆到真核表达载体pRc RSV中 ;用脂质体转染法将此表达载体导入骨髓瘤细胞系SKO 0 0 7和Jurkat细胞中 ,通过点杂交证明其得到有效表达 ;用阻滞电泳法比较野生型gp130和mgp130分子传导IL 6信号的情况。结果 在SKO 0 0 7细胞中IL 6能激活APRF ,而将野生型和突变的gp130cDNA的表达载体分别导入细胞后 ,APRF活性均得到增强 ,但野生型gp130增强的程度较突变体为高 ;在Jur kat细胞中IL 6不能激活核因子 ,导入野生型和突变的gp130cDNA的表达载体后 ,IL 6能激活APRF ,但不能激活NF IL6。与SKO 0 0 7细胞的结果类似 ,野生型gp130激活的信号强于突变体。结论 只含胞外CBD区的mgp130 ,尽管仍能传导IL 6信号 ,但其效率远低于野生型gp130 ,因此gp130的免疫球蛋白样区在IL 6转导中也起着十分重要的作用。
Objective To investigate whether the membrane anchored gp130 mutant containing the second and the third fibronectin type Ⅲ modules of the extracellular region could still transduce IL 6 signal. Method The membrane anchored gp130 mutant cDNA was amplified by the overlap extension PCR and inserted into the mammalian expression vector pRc/RSV. The mutant was proved to be effectively expressed in SKO 007 cells and Jurkat cells by dot blot hybridization. The mutant's effects on IL 6 signaling in these two kinds of cells were detected by electrophoresis mobility shift assay(EMSA). Results In SKO 007 cells, the mutant could enhance the activated APRF, but the effect was not as strong as that of wide type gp130. IL 6 could not activate nuclear factors in Jurkat cells. After the expression vectors pRc/RSVgp130 and pRc/RSV were transfected into Jurkat cells, IL 6 could activate APRF, but could not do NF IL6. Similar to the results in SKO 007 cells, the effects of the mutant was also weaker than that of wild type gp130. Conclusion Although the mutant could still transduce IL 6 signal, it is not as efficiently as the wild type gp130. It suggested that the Ig like domain of gp130 also played an important role in IL 6 signal transduction.
出处
《中华血液学杂志》
CAS
CSCD
北大核心
2000年第12期632-635,共4页
Chinese Journal of Hematology
