前列腺素E2对人肝癌细胞β1-integrin表达调控及相关机制的研究
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国家自然科学基金(81172003,81101496);教育部高等学校博士学科点专项科研基金(20113234120009);江苏高校优势学科建设工程项目(JX10131801021)


The role of PGE2 on β1-integrin expression and its related mechanism in hepatocellular carcinoma
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    摘要:

    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:5 μmol/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),5 μmol/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)。10 μmol/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5 μmol/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。

    Abstract:

    Objective:To investigate the effect of prostaglandin E2 (PGE2) on the expression of β1-integrin and its related signaling pathway in Huh-7 cells. Methods:Huh-7 cells were treated with PGE2,EP1 receptor agonist(17-phenyltrinor Prostaglandin E2,17-PT-PGE2),EP1 receptor antagonist SC19220,nuclear factor-κappaB(NF-κB)inhibitor PDTC. Western blot and immunofluorescence test were employed to detect the expression of β1-integrin and activation of NF-κB in Huh-7 cells. Results:When Huh-7 cells were treated with 5 μmol/L PGE2 for 24 h,the level of β1-integrin was increased by 129.48% (P < 0.01). EP1 receptor agonist (5 μmol/L 17-PT-PGE2) mimicked the effect of PGE2, and increased the expression of β1-integrin by 216.34%(P < 0.01). EP1 receptor antagonist SC19220(10 μmol/L) suppressed PGE2-mediated expression of β1-integrin by 34.51% (P < 0.05). In immunofluorescence assays,NF-κB translocated into the nucleus induced by 17-PT-PGE2 in Huh-7 cells. Further,Western blot assays showed that the level of Phospho-NF-κB-P65 was increased by 209.27%(P < 0.01)after treatment of 5 μmol/L 17-PT-PGE2 for 120 min. NF-κB inhibitor PDTC decreased EP1-mediated expression of β1-integrin by 63.49% (P < 0.01). Conclusion:PGE2 might up-regulate the expression level of β1-integrin through EP1 receptor in Huh-7 cells,which was partly related to the NF-κB signaling pathway.

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王 洁,白小明,张 海,汪亦品,张 丽,马 娟,冷 静.前列腺素E2对人肝癌细胞β1-integrin表达调控及相关机制的研究[J].南京医科大学学报(自然科学版),2013,(3):297-302

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  • 收稿日期:2012-12-11
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  • 在线发布日期: 2013-06-04
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