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目的:阐明前列腺素E2(prostaglandin E2,PGE2)通过EP1受体上调肝癌细胞Huh-7中β1-integrin的表达及其相关的信号转导通路。方法:用PGE2、EP1受体激动剂(17-PT-PGE2)、EP1受体抑制剂SC19220、NF-κB抑制剂PDTC处理Huh-7细胞,通过Western blot、免疫荧光组织化学实验等方法检测β1-integrin蛋白表达水平和NF-κB的活性。结果:5μmol/L的PGE2处理Huh-7细胞24 h后,β1-integrin的表达水平与对照组相比上升了129.48%(P<0.01),5μmol/L EP1受体激动剂17-PT-PGE2处理使细胞β1-integrin的蛋白表达水平升高了216.34%(P<0.01)。10μmol/L的EP1受体抑制剂SC19220处理后β1-integrin表达水平与PGE2组相比下降了34.51%(P<0.05)。免疫荧光组织化学实验显示17-PT-PGE2处理Huh-7细胞120 min后,NF-κB核表达水平明显增加;Western blot实验显示5μmol/L 17-PT-PGE2处理Huh-7细胞120 min后,磷酸化NF-κB-p65上升了209.27%(P<0.01)。NF-κB抑制剂PDTC处理Huh-7细胞后β1-integrin蛋白表达水平与EP1受体激动剂组相比降低了63.49%(P<0.01)。结论:PGE2可通过EP1受体上调Huh-7细胞中β1-integrin的表达,此调节作用可能与NF-κB信号转导通路有关。
Objective: To investigate the effect of prostaglandin E2 (PGE2) on the expression of β1-integrin in Huh-7 hepatoma cells via EP1 receptor and its related signal transduction pathway. Methods: Huh-7 cells were treated with PGE2, EP1 receptor agonist (17-PT-PGE2), EP1 receptor inhibitor SC19220 and NF-κB inhibitor PDTC. Western blot and immunofluorescence staining were used to detect the expression of β1 -integrin protein expression and NF-κB activity. Results: The expression of β1-integrin in Huh-7 cells treated with 5μmol / L PGE2 for 24 h increased by 129.48% (P <0.01) compared with the control group, while the expression of 5μmol / L EP1 receptor agonist 17-PT-PGE2 The protein level of β1-integrin increased by 216.34% (P <0.01). Compared with PGE2 group, the expression level of β1-integrin decreased by 34.51% (P <0.05) after treatment with 10μmol / L EP1 receptor inhibitor SC19220. Immunofluorescence histochemical staining showed that NF-κB nuclear expression was significantly increased after treated with 17-PT-PGE2 for 120 min in Huh-7 cells. Western blot showed that after Huh-7 cells were treated with 5 μmol / L 17-PT-PGE2 for 120 min, Phosphorylation of NF-κB-p65 increased by 209.27% (P <0.01). The expression of β1-integrin protein in Huh-7 cells treated with NF-κB inhibitor PDTC was decreased by 63.49% (P <0.01) compared with EP1 receptor agonist group. Conclusion: PGE2 can up-regulate the expression of β1-integrin in Huh-7 cells through EP1 receptor, which may be related to NF-κB signal transduction pathway.