MEK/ERK信号通路对人结膜上皮细胞增殖的影响及其机制研究

目的:探讨MEK/ERK信号通路对人结膜上皮细胞增殖的影响及其可能的机制。方法:采用不同浓度(0、12.5、25、50、100μmol/L)的MEK抑制剂PD98059处理人结膜上皮细胞(HConEpiC),通过CCK-8法检测不同浓度PD98059作用不同时间(0、12、24、48 h)对人结膜上皮细胞增殖的影响,Western blot检测不同浓度PD98059对人结膜上皮细胞ERK1/2、P-ERK1/2表达的影响。结果:相比对照组(0μmol/L),不同浓度(12.5、25、50、100μmol/L)PD98059处理后的人结膜上皮细胞增殖率明显下降,呈剂量-效应关系,且随处理时间增加(12、24、48 h)其抑制作用也显著增强,差异均有统计学意义(P0.05)。不同浓度PD98059处理人结膜上皮细胞24 h后,其ERK及p-ERK1/2表达随处理浓度增加而降低,与对照组(0μmol/L)相比差异有统计学意义(P0.05),且二者表达量与细胞增值抑制率均呈显著负相关(r=-0.995、r=-0.968,P0.05)。结论:PD98059可抑制人结膜上皮细胞增殖,这可能与其下调ERK表达和减少其活化有关。

Effects and Mechanisms of MEK/ERK Signaling Pathway on the Proliferation of HConEpiC

Objective:To explore the effects and possible mechanisms of MEK/ERK signaling pathway on the proliferation of human conjunctival epithelial cells.Methods:Human conjunctival epithelial cells (HconEpiC) were treated with different concentrations (0, 12.5, 25, 50, 100 umol/L) of MEK inhibitor PD98059. CCK-8 method was used to detect the effect of different concentrations of PD98059 on the proliferation of HconEpiC treated by different time periods (0, 12, 24, 48 h). Western-blot was used to measure the effect of different concentrations of PD98059 on the expressions of ERK1/2, p-ERK1/2 protein.Results:Compared with the control group, the proliferation of HConEpiC was significantly decreased by different concentrations (12.5, 25, 50, 100umol/L) of PD98059 in a concentrations and time periods(0, 12, 24, 48 h) dependent manner (P<0.05). The ERK1/2 and p-ERK1/2 expressions of HConEpiC treated with different concentrations of PD98059 were reduced (P<0.05), and the expressions of ERK and p-ERK were negatively correlated with the inhibiting rates of HconEpiC proliferation (r=-0.995, r=-0.968, P<0.05).Conclusion:PD98059 could inhibit the proliferation of HConEpiC, which might be associated with the inhibition of ERK expresson and activation.