血清诱导95D细胞中Nm23H1从细胞质转位至细胞膜板状伪足

Serum-induced Membrane Translocation of Nm23H1 in 95D Cells

目的 探索Nm2 3H1(肿瘤转移抑制因子之一)在细胞内的定位与其功能关系。方法 以人肺巨细胞癌95D细胞为模型,在血清饥饿2 4h后加入血清,观察细胞形态变化及Nm2 3H1在细胞内的分布变化。结果 血清刺激可以诱导95D细胞形成板状伪足,细胞免疫荧光实验发现Nm2 3H1主要分布于细胞质,但在血清诱导形成的细胞板状伪足部位出现显著的Nm2 3H1分布。亚细胞组分分离及免疫印迹的实验结果也证实Nm2 3H1在血清刺激下可以转位到细胞膜,并且这种诱导作用是迅速的,在30min左右达到高峰,8h后降至仍然略高于血清诱导前的水平。结论 Nm2 3H1这种特殊的定位可能涉及细胞的运动,为进一步研究Nm2 3H1的功能与作用机制提供了基础。

Purpose To explore the cellular distribut ion of Nm23H1 in human giant-cell lung carcinoma 95D cells. Methods We used Immunofluorescence and Western blotting to analysis the distribution of Nm23H1 in 95D cells treated with serum after se rum starvation for 24 hours. Results Serum stimulation ould induced lamellipodia for mation in 95D cells.Most of Nm23H1 existed in cytosol,but there was significant Nm23H1 staining in new formed lamellipodia.Using subcellular fractionation and W estern blotting,we confirmed that Nm23H1 could be translocated to membrane after s erum stimulation,which happened rapidly,peaked at about 30 minutes,and slightly reduced to above basal level at 8 hours. Conclusions Our results indicated that Nm23H1 transloct ion is associated with the formation of lamellipodia,which may provide a clue to further elucidate the role of Nm23H1 played in tumor suppression.