转录因子Foxo3a高表达对小鼠T淋巴瘤EL-4细胞周期和凋亡的影响  

Effects of Over-expressed Transcription Factor Foxo3a on the Cell Cycle and Apoptosis of Mouse T Leukemia EL-4

作　　者：马晓杰[1] 刘光伟[1] 王娜[1] 崔旻[1] 李景鹏[2] 赵勇[1] 

机构地区：[1]中国科学院动物研究所生物膜与膜生物工程国家重点实验室移植生物学研究组,北京100080 [2]东北农业大学生命科学中心基因部,哈尔滨150080

出　　处：《动物学杂志》2006年第5期25-31,共7页Chinese Journal of Zoology

基　　金：中国科学院引进海外杰出人才百人计划资助项目(No.2003-85);国家杰出青年基金(No.C03020504);国家基础研究计划"973项目"(No.2003CB515501);国家教育部海外归国学者科研基金(No.2005-546);中国科学院创新基金(No.KSCX2-SW-333);中国博士后基金项目(No.2004036300);中科院王宽诚博士后科研基金项目(2005)资助

摘　　要：为了研究转录因子Foxo3a高表达对小鼠T淋巴瘤EL-4细胞周期和凋亡的影响,采用电穿孔法将真核表达载体pEGFP-N1/Foxo3a转染小鼠T淋巴瘤细胞系EL-4细胞,并通过聚合酶链式反应和免疫印迹法分别检测Foxo3a mRNA及蛋白表达。转录因子Foxo3a高表达后,采用细胞计数法绘制其细胞生长曲线;采用荧光显微镜法及流式细胞仪定性和定量观察典型EL-4细胞凋亡形态特征、细胞凋亡百分率及细胞周期变化情况。结果表明,转录因子Foxo3a真核表达质粒pEGFP-N1/Foxo3a经酶切鉴定及测序检测序列正确。转染pEGFP-N1/Foxo3a的小鼠EL-4细胞表达Foxo3a mRNA和蛋白水平显著升高。Foxo3a高表达明显抑制EL-4细胞的增殖能力,并使EL-4细胞发生明显G2期阻滞(P<0.001)。Foxo3a基因高表达后,荧光显微镜可以观察到典型凋亡的细胞形态。同时,EL-4细胞凋亡百分率显著升高(P<0.01)。结果提示,Foxo3a高表达可以有效抑制小鼠T淋巴瘤细胞体外细胞增殖,使细胞周期G2时相阻滞,并具有诱导细胞凋亡的作用。To study the effects of overexpressing transcription factor Foxo3a on cell proliferation and apoptosis in mouse T leukemia EL-4 cell line, we have transfected the eukaryntic expression vector pEGFP-NI/Foxo3a to the mouse T leukemia EL-4 cells by electroporation. The expression of Foxo3a was determined by RT-PCR and Western blot analysis. After overexpression of transcription factor Foxo3a, the cell growth curve was evaluated with a cell count method. The EL-4 cell morphology, cell cycle and apoptosis were detected with fluorescent microscopy and flow cytometry. Results showed that the transcription factor Foxo3a eukaryotic expression plasmid pEGFP-NI/Foxo3a sequence is correct with the method of sequencing identification. The EL-4 cells showed increased Foxo3a expression in mRNA and protein levels after transfection with the plasmid pEGFP-NI/Foxo3a. Moreover, these cells expressing high levels of Foxo3a showed a significant decrease in cell proliferation and an significant increase in G2 phase arrest （ P 〈 0.001） as well as a significant increase in apoptosis （P 〈 0.01）. The data indicate that Foxo3a over-expression significantly inhibits the cell proliferation, induces cell cycle G2 phase arrest, and enhances the occurrence of apoptosis in EL-4 cells in vitro.

关 键 词：FOXO3A EL-4细胞 基因高表达 细胞周期 细胞凋亡 

分 类 号：Q343[生物学—遗传学]