LIF受体胞内功能域调节白血病细胞增殖分化

Cytoplasmic domain of leukemia inhibitory factor receptor regulates proliferation and differentiation of leukemia cells

目的:探讨白血病抑制因子受体(LIFR)浕亚基胞内游离功能域gp190CT3对启动靶细胞内信号转导通路(JAK/STAT3),使白血病细胞HL-60向粒细胞方向分化的作用。方法:采用免疫细胞化学、RT-PCR以及流式细胞术等技术和方法,观测pcDNA3.0-gp190CT3重组质粒稳定转染CHO细胞中靶基因的表达;以及稳定转染的CHO细胞与HL-60细胞共培养后,HL-60细胞的形态学、STAT3磷酸化水平、细胞表面抗原CD15表达的变化。结果:(1)重组质粒转染的CHO细胞表达目的基因gp190CT3,并获得稳定表达目的基因的细胞株;(2)重组质粒转染的CHO细胞与野生型HL-60细胞共培养可使HL-60细胞体积变大,形态不规则,STAT3磷酸化水平和细胞表面抗原CD15表达升高。结论:LIF受体α亚基胞内游离片段表达的功能蛋白gp190CT3能够启动靶细胞内JAK/STAT3信号转导通路,从而发挥其调节白血病细胞增殖分化的效能。

Objective：To investigate the effect of the cytoplasmic domain of leukemia inhibitory factor （LIF） receptor a sub mit, gp190CT3, on activation of the JAK/STAT3 signal transduction pathway and on promotion of leukemia cell HL-60 differentiation into granulocytes. Methods： pcDNA3.0 gpl90CT3 was used to transfect CHO cells. Using immunofluorescent cytochemistry, RT-PCR and flow cytometry techniques, we detected the expression of gp190CT3 gene and protein in the stably-transfected CHO cells. Then the stably-trasfected CHO cells were co-cultured with HL-60 cells and the morphological changes of the HL-60 cells were observed, the levels of STAT3 phosphorylation and the expression of the cell surface antigen CD15 were also determined. Results： Expression of gp190CT3 gene was found in pcDNA3. 0-gp190CT3 transfected CHO cells and gp190CT3-stably-transfect CHO cells were obtained. Compared with the untreated HL-60 cells, the size of the co-cultured HL- 60 cells was increased, the morphology was irregular and the level STAT3 phosphorylation and the expression of CD15 were increased. Conclusion： The LIF receptor a submit gp190CT3 participates in the activation of JAK/STAT3 signal transduction pathway and regulates HL-60 cell differentiation and proliferation.