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LIF受体胞内功能域调节白血病细胞增殖分化
姚云,王静,刘厚奇*
0
(第二军医大学基础部组织胚胎学教研室)
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。
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 Houqi*
(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 HL60 differentiation into granulocytes. Methods: pcDNA3.0gp190CT3 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 ɑ submit gp190CT3 participates in the activation of JAK/STAT3 signal transduction pathway and regulates HL60 cell differentiation and proliferation.
Key words:  leukemia inhibitory factor receptor  cell differentiation  signal transduction