核内M-CSF对NIH3T3细胞内微丝的影响  

The Effect of M-CSF in Nucleus on Microfilaments in NIH3T3 Cells

作　　者：张晓红[1] 唐圣松[2] 赵飞骏[3] 刘月顺[1] 龙治锋[1] 

机构地区：[1]南华大学组织胚胎学教研室,湖南衡阳421001 [2]南华大学药物药理研究所,湖南衡阳421001 [3]南华大学病原生物学研究所,湖南衡阳421001

出　　处：《解剖科学进展》2007年第4期314-316,320,共4页Progress of Anatomical Sciences

基　　金：国家自然科学基金(30270684);湖南省杰出青年基金(02JJYB004);湖南省自然科学基金重点项目(04JJ20006)

摘　　要：目的建立M-CSF核内稳定表达的NIH3T3细胞系,探讨核内M-CSF对细胞骨架的影响。方法经脂质体介导核内定位表达重组体pCMV/M-CSF转染NIH3T3细胞,G418筛选后,用Rt-PCR、免疫细胞化学鉴定其在真核细胞中的表达及定位分布,用考马斯亮蓝染色细胞微丝测定M-CSF进入细胞核后对细胞骨架的影响。结果RT-PCR结果显示转染pCMV/M-CSF的NIH3T3细胞能稳定表达M-CSFmRNA;免疫细胞化学结果显示表达的M-CSF定位于NIH3T3细胞核。考马斯亮蓝染色结果显示转染pCMV/M-CSF的NIH3T3细胞内微丝数量减少,排列紊乱。结论核内M-CSF可引起NIH3T3细胞内微丝数量减少,排列紊乱。Objective To establish a cell line which stably expresses macrophage colony-stimulated factor （M- CSF） in NIH3T3 cells nucleus and explore the effect of nuclear M-CSF on the cytoskeleton in NIH3T3 cells. Methods Using liposome, the pCMV/M-CSF was transfected into NIH3T3 cells and the cells clones were selected with G418. The expression and localization of M-CSF in NIH3T3 cells were verified by RT-PCR and immunocytochemistry. The effect of nuclear M-CSF on cytoskeleton was analyzed by dying with Coomassie brilliant blue. Result After transfecting pCMV/M- CSF into NIN3T3 cells by liposome and screening using G418, the pCMV/M-CSF-transfected NIN3T3 cells can stably express both M-CSF mRNA and protein in nucleus. Less and disorderly-arranged microfilaments were found in pCMV/M- CSF-transfected NIN3T3 cells, compared to un-transfected NIN3T3 cells. Conclusion Nuclear M-CSF gives rise to the less and disorderly-arranged microfilaments in NIH3T3 cells.

关 键 词：巨噬细胞集落刺激因子(M—CSF) 核内定位表达 NIH3T3细胞 微丝 

分 类 号：R730.231[医药卫生—肿瘤]