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目的 明确p2 1ras 细胞外信号调节蛋白激酶 (theextracellularsignal regulatedproteinki nase,ERK)系列是否参与调节Jurkat细胞产生TNF β和IL 2。方法 用负向突变基因p2 1ras(ras17N或ras15A) ,raf 1(raf1～ 130 )及ERK1(erk1K71R)或活性结构p2 1ras(H ras6 1L)和raf(raf2 2W)转染细胞 ,以PMA PHA刺激细胞 ,ELISA法检测细胞因子。结果 PMA PHA刺激细胞产生较正常对照细胞约 2倍的TNF β和IL 2 (P <0 .0 0 1)。负向突变基因去除了PMA PHA增加细胞因子分泌的作用。活性突变基因本身不影响TNF β和IL 2产生 ,但明显增强细胞对PMA PHA的应答 (P <0 .0 5 )。结论 p2 1ras ERK系列在调节Jurkat细胞在PMA PHA刺激下产生TNF β和IL 2中起重要的作用。
Objective To investigate whether the extrinsic signal regulated protein kinase (ERK) of p2 1ras is involved in regulating the production of TNF β and IL 2 in Jurkat cells. Methods The cells were transfected with negative mutant gene p2 1ras (ras17N or ras15A), raf1 (raf1-130) and ERK1 (erk1K71R) or active structure p2 1ras (ras6 1L) and raf2 2W Cell, ELISA to detect cytokines. Results PMA PHA stimulated cells to produce approximately two-fold more TNF [beta] and IL2 than normal control cells (P <0. 001). The negative mutant gene removes the effect of PMA PHA on cytokine secretion. The active mutant gene itself did not affect the production of TNF [beta] and IL2, but significantly enhanced the cell response to PMA PHA (P <0.05). Conclusion The p2 1ras ERK series plays an important role in regulating the production of TNF β and IL 2 by Jurkat cells stimulated by PMA PHA.