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目的 探讨人白血病Meg 0 1细胞的异常增殖机制以及凋亡因子Fas对细胞异常增殖的调节作用。方法 用基因重组技术将Fas的细胞外区、跨膜区与白血病抑制因子 (LIF)受体两亚基(gp190 ,gp130 )细胞内区构成嵌合型受体 (Fas 190 ,Fas 130 ) ,同时 ,将Fas死亡区域 (FASDD)替换Fas 130中gp130细胞内区无结构域部分 (Fas 130f) ,分别在Meg 0 1细胞内表达后 ,用抗Fas抗体激活这些嵌合型受体 ,随后用免疫组化和免疫印迹法分析受体细胞内区形成同源性寡聚体 (190cyt 190cyt 190cyt,130cyt 130cyt 130cyt及FASDD FASDD FASDD)后的细胞状况和细胞内c myc的表达。结果 转染pEDFas 130后用抗Fas抗体作用 6～ 8h ,Meg 0 1细胞c myc表达量增加 ,细胞增殖明显 ;而pEDFas 190形成寡聚体后 ,Meg 0 1细胞c myc的表达有所下降。另外 ,Fas 130与Fas 130f比较 ,后者的c myc的表达明显下降 ,细胞形态大小不一。结论 LIF受体中gp130亚基细胞内区促进白血病Meg 0 1细胞内的c myc表达及细胞增殖信号的传递 ;Fas死亡域可抑制gp130的细胞增殖活性。
Objective To investigate the abnormal proliferation mechanism of human leukemia Meg 0 cells and the regulatory effect of Fas on the abnormal proliferation of human leukemia cells. Methods The chimeric receptors (Fas 190 and Fas 130) were constructed by intracellular domain of the extracellular domain and transmembrane region of Fas with the two subunits of gp190 and gp130 of the leukemia inhibitory factor (LIF) , The Fas death domain (FASDD) was substituted for the non-domain portion of gp130 intracellular domain (Fas130f) in Fas130, respectively, after being expressed in MegOl cells, these chimeric receptors were activated with anti-Fas antibody followed by immunization Histological and immunocytochemical analysis of cellular status and intracellular c-myc expression after formation of homologous oligomers (190cyt 190cyt 190cyt, 130cyt 130cyt 130cyt and FASDD FASDD FASDD) in the intracellular regions of the recipient cells. Results After transfection of pEDFas 130 with anti-Fas antibody for 6 ~ 8 h, the expression of c-myc in Meg 0 1 cells was increased and the cell proliferation was obvious. However, the expression of c-myc in Meg 0 cells decreased after pEDFas 190 oligomer formation. In addition, compared with Fas130f, the expression of cmyc in the latter was significantly decreased and the cell morphology varied. Conclusion The intracellular gp130 subunit of LIF receptor can promote the expression of c-myc in leukemia cells and the signal of cell proliferation in leukemia cells; the death domain of Fas can inhibit the cell proliferation activity of gp130.