摘要
以PC12细胞为神经元细胞模型,研究Nogo-C对神经元细胞存活及增殖的作用。在PC12细胞中转染过表达Nogo-C,使用G418药物筛选以获得稳定表达的细胞克隆,利用Hoechst33342染色、细胞计数、MTT以及流式细胞仪等技术检测Nogo-C对细胞增殖以及细胞周期的影响。结果表明:(1)Hoechst33342染色未观察到表达Nogo-C的细胞发生明显凋亡;(2)细胞计数及MTT实验观察到转染Nogo-C后的PC12细胞生长增殖活性明显降低;(3)流式细胞仪检测细胞生长周期,正常PC12细胞G1期的百分数为(37.8±7.9)%,S期为(50.4±8.5)%,而转染Nogo-C的PC12细胞G1期为(76.8±4.1)%,S期为(14.7±1.7)%,提示转染Nogo-C的PC12细胞的细胞周期被阻滞在G1期;(4)没有获得稳定表达Nogo-C的PC12细胞模型。实验证明,过表达Nogo-C通过使PC12细胞周期被阻滞在G1期而明显抑制细胞的增殖,但是并不引起细胞的凋亡。
The aim was to study the effect of Nogo-C on neuronal survival and proliferation, using PC12 cell as a neuronal model. The cells were transfected and overexpressed with Nogo-C and selected by G418 to obtain stable clones. The effect on cell survival and proliferation was studied by Hoechst 33342 staining, cell counting, MTT assay, and flow cytometry. Our results revealed that： （1） no obvious apoptosis was found in the Nogo-C overexpressed PC12 cells shown by Hoechst 33342 staining; （2） cell proliferation was inhibited significantly in the Nogo-C overexpressed PC12 cells, as detected by cell counting and MTT assay： （3） FACS analysis demonstrated that in the normal control the cells at the G1 and S phases were （37.8±7.9）%, （50.4±8.5）% respectively, whereas the Nogo-C overexpressed cells at the G1 and S phases were （76.8±4.1）% and （14.7±1.7）% respectively, indicating that the arrest of cell cycle of the Nogo-C overexpressed cells occurred at phase G1; （4） we were not able to obtain stable clones of Nogo-C overexpressed PC12 ceils, In conclusion, overexpression of Nogo-C in PC12 cells inhibits cell proliferation through impeding cell cycle at phase G1, but do not induce apoptosis.
出处
《细胞生物学杂志》
CSCD
2006年第4期582-586,共5页
Chinese Journal of Cell Biology
基金
国家重点基础研究发展规划(973计划)资助项目(No.2003CB515301)~~
