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探讨人白血病细胞系U937白血病抑制因子 (LIF)受体α亚基和另一亚基gp130细胞内区与促分裂原活化蛋白激酶 (MAPK)的关系 ,旨在研究白血病细胞增殖和分化的机制。用基因重组技术将两基因细胞内区互换以构成两嵌合体受体 (190 130 ,130 190 )并分别在U937表达 ,其与野生受体竞争性结合白血病抑制因子 ,用免疫组化和免疫印迹法分析受体细胞内区形成同源性二聚体(190cyt 190cyt,130cyt 130cyt)后的细胞状况和细胞内MAPK的水平。结果表明 ,转染pE190 130后用LIF作用 6h ,U937细胞MAPK表达量增加 ,MAPK形成的二聚体较明显 ,细胞增殖较快 ;而另一嵌合体受体与α亚基形成 190cyt 190cyt时U937细胞MAPK的表达无变化 ,二聚体不明显。说明LIF受体中gp130亚基的细胞内区参与了MAPK的激活及白血病U937细胞增殖信号的传递。
To investigate the relationship between the intracellular domain of human leukemia cell line U937 leukemia inhibitory factor (LIF) receptor alpha and another subunit of gp130 and mitogen-activated protein kinase (MAPK) in order to study the mechanism of leukemic cell proliferation and differentiation. The intracellular regions of both genes were interchanged by genetic recombination techniques to form two chimeric receptors (190 130, 130 190) and expressed separately at U937, which competitively bound leukemia inhibitory factor with wild-type receptors, with immunohistochemistry and immunization The cell morphology and the level of intracellular MAPK after the formation of homodimers (190cyt 190cyt, 130cyt 130cyt) were analyzed by Western blotting. The results showed that the expression of MAPK in U937 cells increased after transfection of pE190 130 with LIF for 6h, the dimer formed by MAPK was more obvious, and the cell proliferation was faster. When the other chimera receptor and α subunit formed 190cyt 190cyt, U937 Cell MAPK expression did not change, dimer is not obvious. This shows that the intracellular domain of gp130 subunit of LIF receptor is involved in the activation of MAPK and the signal of proliferation of leukemia U937 cells.