PD98059对肝癌HepG2细胞Tec信号转导作用的初步研究

Effect of MEK1 inhibitor PD98059 on the signal transduction of Tec in HepG2 hepatoma cells

目的探讨MEK1抑制剂PD98059对肝癌细胞株HepG2细胞中Tec(tyrosinekinaseexpressedinhepatocellu-larcarcinoma)及ERK2作用。方法免疫细胞化学方法检测Tec、ERK2在HepG2细胞中的表达;用RT-PCR和Westernblot方法检测,不同浓度PD98059处理细胞后,HepG2细胞中的Tec、ERK2mRNA及蛋白表达。结果Tec、ERK2在HepG2细胞中呈高表达,PD98059明显影响HepG2细胞Tec、ERK2mRNA以及蛋白表达,呈剂量依赖性,40μmol/LPD98059细胞抑制最明显。结论Tec可能是肝癌细胞内Ras/Raf/ERK信号转导途径上游的信号蛋白,与Ras/Raf/ERK信号转导通路存在着某种信号联系。

Objective To investigate the effect of MEK1 inhibitor PD98058 on Tee and ERK2 in HepG2 hepatoma cells. Methods The expression of mRNA and protein of Tec and ERK2 in HepG2 cells was detected by immunocytochemistry assay. After various concentration of PD98059 treatment, the expression of Tec and ERK2 mRNA in HepG2 cells was detected by RT-PCR and Western blotting. Results Tec and ERK2 expressed highly in HepG2 cells. PD98059 obviously inhibited the expression of mRNA and protein of Tec and ERK2 in a dose-dependent manner, in which 40 μ mol/L of PD98059 exhibited the strongest inhibiting effect. Conclusion PD98058, as MEK1 inhibitor, can inhibit Tec, block the signal route of Ras/Raf/ERK and to impede the signal transduction in HepG2 cells. Tec may be the signal protein in the upper stream of Ras/Raf/ ERK in hepatocarcinoma cells and is supposed to interact with the signal way of Ras/Raf/ERK.