维甲酸促进小鼠F9细胞中STAT1的反式激活作用

Retinoic Acid Stimulates the Transactivation of STAT1 in Mouse F9 Cells

维甲酸能促进肿瘤细胞的凋亡,诱导体外胚胎干细胞的分化,但作用机制的不明使其应用受到极大限制.因此,更多更全面地了解维甲酸的作用机制具有重要意义.本文构建了信号传导与转录激活因子(STAT1和STAT3)的真核表达载体,通过免疫荧光染色、电泳迁移率实验以及双荧光素酶报告基因检测系统证明,维甲酸诱导可以激活转录因子STAT1促使其进入细胞核,并且增强STAT1蛋白与靶基因启动子的结合能力,从而发挥基因表达调控作用.本文结合后续的STAT1功能分析,试图建立起一种"维甲酸-转录因子-靶基因"的研究模式,有助于维甲酸作用机制的全面、系统的研究.这为临床上使用维甲酸作为抗肿瘤药提供理论基础,同时也为胚胎干细胞多能性调控机制研究提供新思路.

Retinoic acid induces the apoptosis of tumor cells,the differentiation of embryonic stem cell in vitro,but how did it works was not completely known,which imposes restrictions on its clinical application.Hence studies on the mechanism of retinoic acid will be of great significance.Toward this end,we constructed eukaryotic expression plasmid of STAT1 and STAT3,transient infection and Western blotting indicated that the plasmids can be efficiently expressed in F9 embryonal carcinoma cells.Further experiments using immunofluorescence,electrophoretic mobility shift assay（EMSA） and dual luciferase reporter（DLR） assay proved that retinoic acid treatment stimulates the transactivation of STAT1 by translocating it to nucleus and enhancing its DNA binding ability.This work followed by the study of STAT1 function could form a ＂retinoic acid-transcription factor-target gene＂ model,it will make a contribution to the mechanism research of retinoic acid.The results provide a theoretical principle for retinoic acid as an antineoplastic drug,as well as put new insight to the research of pluripotency regulation of embryonic stem cells.