EP1受体介导PGE2对胆管细胞癌HuCCT1细胞MMP2表达和活性的影响
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国家自然科学基金资助(30871015,30900583)


Prostaglandin E2 upregulates MMP2 expression and activity in HuCCT1cells via EP1 receptor signaling pathway
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    摘要:

    目的:探讨前列腺素E2(PGE2)对人胆管细胞癌HuCCT1细胞基质金属蛋白酶2(MMP2)的表达与酶活性的影响及与EP1受体的关系-方法:分别用PGE2-EP1受体激动剂或抑制剂-蛋白激酶C(PKC)抑制剂和钙离子螯合剂处理HuCCT1细胞,通过RT-PCR-明胶酶谱法检测MMP2 的mRNA水平以及MMP2酶活性-结果:5 μmol/L PGE2和5 μmol/L EP1受体激动剂17-PT-PGE2处理组MMP2 mRNA的表达水平与对照组相比分别上升了89.14%(P < 0.01)和163.89%(P < 0.01);MMP2酶活性与对照组相比分别增强了69.11%(P < 0.05)和117.65%(P < 0.01);10 μmol/L EP1受体抑制剂sc-51322处理后,MMP2 mRNA水平以及MMP2酶活性较PGE2处理组分别下降了47.74%(P < 0.05)-84.58%(P < 0.01);5 μmol/L PGE2或5 μmol/L 17-PT-PGE2处理稳定转染EP1R-pcDNA3的人胚肾细胞株HEK293细胞,MMP2的酶活性较对照组分别增强了36.07%(P < 0.05)-61.59%(P < 0.05)-5 μmol/L PKC抑制剂BIS-1-10 μmol/L钙离子螯合剂BAPTA-AM处理后,MMP2 mRNA水平较17-PT-PGE2处理组分别下降了44.17%(P < 0.05)-34.42%(P < 0.05);MMP2酶活性较17-PT-PGE2处理组分别下降了70.95%(P < 0.05)-71.82%(P < 0.05)-结论:PGE2 可以通过EP1受体上调胆管细胞癌HuCCT细胞MMP2 mRNA的表达以及MMP2的酶活性,此调节作用可能与Ca2+/PKC信号转导通路有关-

    Abstract:

    Objective:To investigate the effect of prostaglandin E2 (PGE2) on expression and enzyme activity of matrix metalloproteinase 2(MMP2) by EP1 receptor in cholangiocarcinoma cells HuCCT1 cells. Methods:HuCCT1 cells were treated with PGE2, EP1 receptor agonist, EP1 receptor antagonist, protein kinase C(PKC) inhibitor and Ca2+ chelating agent, RT-PCR and gelatin zymography were employed to detect the mRNA level and enzyme activity of MMP2 in HuCCT1 cells. Results:The mRNA levels of MMP2 increased 89.14%(P < 0.01), 163.89%(P < 0.01); and the enzyme activity of MMP2 increased 69.11%(P < 0.05), 117.65(P < 0.01) compared with the control group after treated with 5 μmol/L PGE2, 5 μmol/L EP1 receptor agonist 17-PT-PGE2 respectively. The mRNA level and the enzyme activity of MMP2 in HuCCT1 cells treated with 10 μmol/L EP1 receptor antagonist sc-51322 decreased 47.74%(P < 0.05),84.58%(P < 0.01) compared with the cells treated with PGE2. After treated with 5 μmol/L PGE2 or 5μmol/L 17-PT-PGE2, the enzyme activity of MMP2 in EP1R-pcDNA3 transfected HEK293 cells increased 36.07%(P < 0.05), 61.59%(P < 0.05) compared with the control group. When treated with 5 μmol/L PKC inhibitor BIS-1, 10 μmol/L Ca2+ chelating agent BAPTA-AM, the mRNA levels of MMP2 decreased 44.17%(P < 0.05),34.42%(P < 0.05); and the enzyme activities of MMP2 decreased 70.95%(P < 0.05),71.82%(P < 0.05) compared with the cells treated with 17-PT-PGE2. Conclusion:PGE2 might up-regulate the mRNA level and the enzyme activity of MMP2 through EP1 receptor in cholangiocarcinoma HuCCT1 cells, which could be partly related to the Ca2+/PKC signaling pathway.

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汪亦品,张 丽,束 为,孙 波,张 海,白小明,彭 韬,冷 静. EP1受体介导PGE2对胆管细胞癌HuCCT1细胞MMP2表达和活性的影响[J].南京医科大学学报(自然科学版),2011,(7):930-934

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  • 收稿日期:2011-01-24
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