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目的研究丝裂原活化蛋白激酶(MAPK)亚族细胞外调节蛋白激酶(ERK)通路在HepG2肝癌细胞迁移和侵袭中的作用。方法体外培养人肝癌HepG2细胞,采用ERK通路激动剂和抑制剂:表皮生长因子(EGF)及U0126刺激48h,分别应用MTS法、划痕实验法及Transwell小室法检测EGF和U0126对HepG2细胞增殖能力、迁移能力和侵袭能力的影响,qPCR和Western blot法分别检测ERK mRNA和磷酸化ERK蛋白的表达变化。结果细胞增殖、迁移和侵袭实验结果均显示,激动剂EGF可显著提高HepG2肝癌细胞的增殖、迁移及侵袭能力(P<0.01),而U0126均对以上指标有抑制效应(P<0.05)。qPCR和Western blot结果显示,EGF可上调ERK mRNA和磷酸化蛋白的表达(P<0.01),而U0126对ERK mRNA无明显影响(P>0.05),但可显著下调ERK磷酸化蛋白的表达(P<0.01)。结论 ERK通路参与HepG2细胞的增殖、迁移和侵袭过程,EGF和U0126对以上指标的影响可能与ERK mRNA的表达及蛋白磷酸化过程相关。
AIM To investigate the role of mitogen-activated protein kinase (MAPK) sub-family of extracellular regulatory protein kinase (ERK) pathway in the migration and invasion of HepG2 hepatocarcinoma cells. Methods HepG2 cells were cultured in vitro and stimulated by EGF and U0126 for 48h. MTS assay, scratch assay and Transwell chamber assay were used to detect the proliferation of HepG2 cells , Migration ability and invasion ability, qPCR and Western blot were detected ERK mRNA and phosphorylated ERK protein expression changes. Results The results of cell proliferation, migration and invasion showed that EGF could significantly increase the proliferation, migration and invasion ability of HepG2 hepatocarcinoma cells (P <0.01), while U0126 had inhibitory effect on the above parameters (P <0.05). qPCR and Western blot showed that EGF upregulated the expression of ERK mRNA and phosphorylated protein (P <0.01), while U0126 had no significant effect on ERK mRNA (P> 0.05), but downregulated the expression of ERK phosphorylation protein <0.01). Conclusion The ERK pathway is involved in the proliferation, migration and invasion of HepG2 cells. The effects of EGF and U0126 on the above indexes may be related to the ERK mRNA expression and protein phosphorylation.