摘要: |
目的:探讨白血病抑制因子受体(LIFR)ɑ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HL60向粒细胞方向分化的作用。 方法:采用免疫细胞化学、RTPCR以及流式细胞术等技术和方法,观测pcDNA3.0gp190CT3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HL60细胞共培养后,HL60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HL60细胞共培养可使HL60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。 |
关键词: 白血病 白血病抑制因子受体 细胞分化 信号转导 |
DOI:10.3724/SP.J.1008.2007.01165 |
投稿时间:2007-07-03修订日期:2007-09-25 |
基金项目:上海市科学委员会登山计划(064119612) |
|
Cytoplasmic domain of leukemia inhibitory factor receptor regulates proliferation and differentiation of leukemia cells |
YAO Yun,WANG Jing,LIU Houqi* |
(Department of Embryology and Histology, College of Basic Medical Sciences,Research Center of Developmental Biology, Second Military Medical University, Shanghai 200433,China) |
Abstract: |
Objective:To investigate the effect of the cytoplasmic domain of leukemia inhibitory factor (LIF) receptor ɑ submit, gp190CT3, on activation of the JAK/STAT3 signal transduction pathway and on promotion of leukemia cell HL60 differentiation into granulocytes. Methods: pcDNA3.0gp190CT3 was used to transfect CHO cells. Using immunofluorescent cytochemistry, RTPCR and flow cytometry techniques, we detected the expression of gp190CT3 gene and protein in the stablytransfected CHO cells. Then the stablytrasfected CHO cells were cocultured with HL60 cells and the morphological changes of the HL60 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 pcDNA3.0gp190CT3 transfected CHO cells and gp190CT3stablytransfect CHO cells were obtained. Compared with the untreated HL60 cells, the size of the cocultured HL60 cells was increased, the morphology was irregular and the level STAT3 phosphorylation and the expression of CD15 were increased. Conclusion: The LIF receptor ɑ submit gp190CT3 participates in the activation of JAK/STAT3 signal transduction pathway and regulates HL60 cell differentiation and proliferation. |
Key words: leukemia inhibitory factor receptor cell differentiation signal transduction |