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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:5μmol/L PGE2和5μmol/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);10μmol/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);5μmol/L PGE2或5μmol/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)。5μmol/L PKC抑制剂BIS-1、10μmol/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。
Objective: To investigate the effect of prostaglandin E2 (PGE2) on the expression and enzyme activity of matrix metalloproteinase 2 (MMP2) in human cholangiocarcinoma HuCCT1 cells and its relationship with EP1 receptor. Methods: HuCCT1 cells were treated with PGE2, EP1 agonist or inhibitor, protein kinase C (PKC) inhibitor and calcium chelator respectively. The MMP2 mRNA and MMP2 activity were detected by RT-PCR and gelatin zymography . Results: Compared with the control group, the expression of MMP2 mRNA increased by 89.14% (P <0.01) and 163.89% (P <0.01) respectively in 5μmol / L PGE2 and 5μmol / L EP1 receptor agonist 17-PT- ; MMP2 activity increased by 69.11% (P <0.05) and 117.65% (P <0.01) respectively compared with the control group; MMP-2 mRNA level and MMP2 activity after 10μmol / L EP1 receptor inhibitor sc-51322 treatment PGE2 treatment group were decreased by 47.74% (P <0.05), 84.58% (P <0.01); 5μmol / L PGE2 or 5μmol / L 17-PT-PGE2 treatment of EP1R-pcDNA3 stably transfected human embryonic kidney cell line HEK293 cells , MMP2 increased 36.07% (P <0.05) and 61.59% (P <0.05) respectively compared with the control group. Compared with 17-PT-PGE2 treatment group, MMP-2 mRNA expression decreased by 44.17% (P <0.05) and 34.42% (P <0.05) after 5μmol / L PKC inhibitor BIS-1 and 10μmol / L calcium chelator BAPTA- 0.05). The activity of MMP2 was decreased by 70.95% (P <0.05) and 71.82% (P <0.05), respectively, compared with 17-PT-PGE2 treatment group. CONCLUSION: PGE2 can up-regulate the expression of MMP2 mRNA and the activity of MMP2 in HuCCT cells via EP1 receptor, which may be related to Ca2 + / PKC signal transduction pathway.