p27kip1和Spy1在肝癌细胞HuH7增殖过程中的表达变化及其相互关系

目的探讨p27kip1和Spy1在肝癌细胞HuH7增殖过程中的表达变化及相互关系。方法采用血清饥饿合并释放处理肝癌细胞株HuH7，通过流式细胞术检测该处理对HuH7细胞生长周期的影响；同时采用Western blot、免疫荧光技术检测不同生长状态的HuH7细胞中p27kip1和Spy1的表达及亚细胞定位情况；采用免疫沉淀方法检测HuH7细胞中p27kip1与Spy1的结合情况。结果血清饥饿造成HuH7细胞生长周期停滞，Spy1表达下降，p27kip1蛋白总量增加，差异有统计学意义（P〈0．05或〈0．01）。与此同时，p27kip1的阳性信号也由整个细胞分布转变为胞核高分布，而Spy1的阳性信号则由胞核高分布转变为胞浆高分布。血清释放后刺激细胞进入细胞周期，上述分子呈现相反的变化，且p27州的阳性信号在胞浆分布增强。结论Spy1可能通过与p27kip1结合来介导p27kip1的核内外分布并影响其表达，从而参与调控肝癌细胞的生长。

Expression and Relationship of p27kipl and Spy1 During the Proliferation Process of Hepatocellular Carcinoma Cell HuH7

Objective To investigate the expression and relationship of p27kip1and Spyl during proliferation process of Hepatocellular carcinoma cell HUH7. Methods HuH7 cells were treated with serum starvation and release, and effect on the cell growth with these treatment was tested with flow ey- tometry. The expression and localization of p27kip1 and Spyl in HuH7 cell in different growth condition were detected by Western blot and immunofluorescence. The combination of p27kip1 and Spyl was de- tected with immunopreeipitation. Results The cell cycle of HuH7 was blocked by serum starvation. The expression of Spyl was down-regulated. The albumen amount of p27kip1 increased. Meanwhile, the location of p27kip1 was accumulated in nuclei. However, the location of Spyl was transferred from nuclei to cytoplasm .The HuH7 ceils were reentering the cell cycle after serum release and the reverse changes of these proteinum happened and the location of p27kip1 in cytoplasm was reinforced. Conclusion Spyl may impact the location and expression of p27kip1 through integrating with p27kip1, and then may partici- pate in regulating the growth of Hepatocellular carcinoma cell.