GAM cDNA稳定转染的M1细胞系的建立与鉴定  

Establishment and verificatin of GAM cDNA stably transfected M1 cell line

作　　者：刘飞[1] 刘崟[1] 金伯泉[1] 朱勇[1] 陈丽华[1] 张建平[1] 

机构地区：[1]第四军医大学免疫学教研室,陕西西安710032

出　　处：《免疫学杂志》2000年第3期179-181,185,共4页Immunological Journal

基　　金：国家自然科学基金资助项目!(39700073)

摘　　要：目的为搞清 GAM在 gp130介导的信号传导中所起的作用。方法选择在 IL - 6刺激下出现分化和增殖抑制的M1细胞系 ,分别用正向和反向插入 GAM c DNA的真核表达载体 p EF- BOS与 p SV2 - neo质粒共转染 ,经 G418选择培养筛选得到稳定转染的细胞 ,并用半套式 PCR方法证实了 GAM基因与这两种细胞基因组 DNA的整合。结果流式细胞仪分析表明正向 GAM c DNA转染的 M1细胞中 GAM的表达水平明显升高 ,3H掺入实验表明 GAM表达水平升高使 M1细胞对 IL - 6的反应性下降。结论 GAM高表达 M1细胞的建立为深入研究 GAM在ObjectiveTo investigate the role of GAM gp130 associated molecule in the gp130 mediated signaling.Me thods Under the stimulation of IL6 and sIL6R, whose signal is transducted by gp130, M1 cells, a murine myeloid leukemia cell line, go into growth arrest To investigate the role of GAM in the gp130 mediated signaling, the mammalian expressing vector pEFBOS containing GAM cDNA or antisense cDNA of GAM was constructed and transfected permanently into the M1 cells NestPCR and flow cytometric analysis verified the stable transfectants and increased level of GAMResulstThe increase of GAM expression inhibited the growth arrest effect of IL6 on M1 cells significantly, while the decreasing of GAM enhanced this effect slightlyConclusionThese results indicated that GAM might play a role of negative regulator in gp130 signaling [

关 键 词：gp130结合分子 信号传导 基因转染 GAM CDNA 

分 类 号：R392.11[医药卫生—免疫学]