摘要
目的 探讨蛋白激酶Cα(PKCα)对人正常肝和肝癌细胞周期的作用和对G1 期相关调控因子的影响。 方法 通过细胞转染技术将PKCαcDNA正向插入的真核表达质粒PXJ4 1 PKCα导入正常肝细胞 (L 0 2 ) ,并利用本室已构建的表达反义PKCα的BEL 74 0 2细胞 (HT6 )检测细胞的生长曲线 ,细胞周期以及细胞对G1 期相关调控因子cylinD1和cyclinE的影响。 结果 构建了稳定过表达PKCα的人正常肝细胞模型 (LT3) ,过表达PKCα可促进L 0 2细胞增殖 ,促进细胞由G1 期向S期的过渡 ;cyclinD1和cyclinE的蛋白水平上升 ,反之表达反义PKCα的BEL 74 0 2细胞 (HT6 )增殖被抑制 ,阻抑细胞由G1 期向S期的过渡 ,cyclinD1和cyclinE的蛋白水平下降。 结论 从正反两个方面表明 ,PKCα可通过作用于G1 期相关周期蛋白的水平影响G1 S期的进程。
Objective To study the effect of protein kinase Cα(PKCα) on cell cycle and G 1 phase related regulatory protein——cyclinD1 and cyclinE in human normal liver cells and hepatoma cells in vitro. Methods The plasmid PXJ41-neo and PXJ41-PKCα were thansfected into human normal liver cells and characterized by RT-PCR and Western blotting, the growth curve, cell cycle and the effect on G 1 phase related regulatory protein——cyclinD1 and cyclinE were tested by FCM and Western blotting respectively. Results The cell model(LT3)overexpressing PKCα was constructed.It was found that the growth and progression of G 1/S were promoted in transfected human normal liver cells (LT3), the level of cyclinD1 and cyclinE was increased. On the contrary, The growth and progression of G 1/S were decreased in human hepatoma cells expressing antisense PKCα(HT6),the level of cyclinD1 and cyclinE was inhibited.Conclusion The progression of G 1/S is effected by G 1 phase related regulatory protein——cyclinD1 and cyclinE, which is regulated by PKCα in human normal liver cells and hepatoma cells.
出处
《解剖学报》
CAS
CSCD
北大核心
2005年第1期37-40,共4页
Acta Anatomica Sinica
基金
国家自然科学基金资助项目 (3 9870 866)
