巨噬细胞移动抑制因子（MIF）调节细胞凋亡、细胞生长及周期的分子机制研究

【作者】 刘凌峰；

【导师】 谢毅；

【作者基本信息】 复旦大学 ， 遗传学， 2008， 博士

【摘要】 本论文研究通过酵母双杂交技术首次发现MIF与Bim相互结合,并通过invitro和in vivo蛋白结合技术证实了MIF和Bim的相互作用,并通过不同分子生物学技术证实了MIF参与反向调控Bim介导的线粒体凋亡途径,为内源性MIF提高细胞抗凋亡的能力提供了新的分子依据,并为研究线粒体凋亡途径中Bim的调节机制提供了新的思路。在研究MIF调节Bim介导的细胞凋亡的过程中,发现在HEK293细胞内抑制MIF表达后,细胞增殖活力降低,通过流式细胞仪细胞周期检测证实,MIF knockdown可阻碍细胞周期进程,导致细胞停滞于G0/G1期:为了阐明MIF调控细胞周期潜在的分子机制,我们应用microarray技术分析了MIF(+)和MIF(-)细胞的全基因组的mRNA表达谱差异,并通过细胞内信号通路分析方法结合MIF相关的基因差异性表达谱的数据,首次建立在细胞周期G1/S期进程中,以MIF knockdown为原点的信号通路调控网络,生动再现了在细胞内MIF影响细胞生长和细胞周期进程的动态的信号转导通路,这为揭示MIF在细胞内的分子生物学功能提供了有力的线索。更多还原

【Abstract】 In the first section of the dissertation,we have employed a yeast two-hybrid system to isolate a novel Bim-interacting protein,macrophage migration inhibitory factor(MIF).The interaction between MIF and Bim was confirmed by both in vitro and in vivo protein interaction assays.Moreover,MIF may regulate pro-apoptotic activity of Bim and inhibit the release of cytochrome c from mitochondria to the cytosol during apoptosis.This study provides the novel insights into the MIF’s anti-apopotic activity and Bim-mediated mitochondria-dependent apoptosis.In the second section of the dissertation,we demonstrate siRNA-mediated knockdown of MIF results in inhibition of cell proliferation and G0/G1 cell cycle arrest in HEK293 cells.To elucidate the molecular mechanism of cell cycle perturbation following MIF knockdown, we employed microarray to investigate the genome-wide expression profile in MIF-deficient cells and normal cells.The dynamic model of MIF-dependent signaling pathways linked to cell cycle machinery were constructed through analyzing gene expression data in the context of the gene ontology and the known biological pathways. We have established an integrate and dynamic signaling network in the MIF knockdown cell.This study provides novel insights into the pleiotropic activities of endogenous MIF, especially its essential and crucial role in cell proliferation and the cell cycle.更多还原