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目的 :探讨前列腺素E2(prostaglandin E2,PGE2)通过EP2受体影响人胆管细胞癌细胞HuCCT1细胞间黏附因子-1(intercellular adhesion molecule-1,ICAM-1)的表达及癌细胞的侵袭能力。方法:用PGE2、EP1～4四种受体激动剂(17-phenyltrinorProstaglandin E2、Butaprost、Sulprostone和Prostaglandin E1 Alcohol)、EP2受体抑制剂AH6809、腺苷酸环化酶(AC)抑制剂SQ22536和蛋白激酶A(PKA)抑制剂H89处理HuCCT1细胞,通过Western blot、划痕试验等方法检测ICAM-1的蛋白表达水平以及HuCCT1细胞侵袭能力的变化。结果:PGE2明显提高HuCCT1细胞ICAM-1蛋白的表达水平,5μmol/L PGE2处理HuCCT1细胞24 h后,ICAM-1蛋白表达水平与对照组相比上升了66.17%(P<0.05),并呈浓度依赖性和时间依赖性;5μmol/L PGE2处理HuCCT1细胞24 h后,HuCCT1细胞的侵袭能力较对照组增强了43.29%(P<0.01)。10μmol/L EP1～4受体激动剂处理HuCCT1细胞24 h后,ICAM-1蛋白的表达水平与对照组相比明显增高,其中EP2受体激动剂上升了257.88%(P<0.05),10μmol/L EP2受体激动剂处理HuCCT1细胞24 h后,HuCCT1细胞的侵袭能力较对照组增强了56.99%(P<0.01);10μmol/LEP2受体抑制剂AH6809处理后ICAM-1蛋白的表达水平与PGE2组相比下降了49.14%(P<0.05),细胞侵袭能力下降了52.06%(P<0.01)。25μmol/L AC抑制剂SQ22536、10μmol/L PKA抑制剂H89处理HuCCT1细胞后,ICAM-1蛋白的表达水平较EP2受体激动剂处理组分别下降了72.87%(P<0.05)和80.78%(P<0.05)。结论:PGE2可通过EP2受体激活cAMP-PKA信号转导通路上调HuCCT1细胞ICAM-1的表达,从而促进HuCCT1细胞的侵袭转移。
Objective: To investigate the effect of prostaglandin E2 (PGE2) on the expression of intercellular adhesion molecule-1 (ICAM-1) and the invasion ability of HuCCT1 cells by EP2 receptor. Methods: PGE2, EP1 ~ 4 receptor agonists (17-phenyltrinorProstaglandin E2, Butaprost, Sulprostone and Prostaglandin E1 Alcohol), EP2 receptor inhibitor AH6809, adenylate cyclase (AC) inhibitor SQ22536 and protein HuCCT1 cells were treated with the inhibitor of kinase A (PKA) H89. The protein expression of ICAM-1 and the invasion ability of HuCCT1 cells were detected by Western blot and scratch assay. Results: PGE2 significantly increased the ICAM-1 protein expression in HuCCT1 cells. After treated with 5μmol / L PGE2 for 24 h, the ICAM-1 protein expression increased by 66.17% (P <0.05) compared with the control group, Dependent and time-dependent manner. The invasive ability of HuCCT1 cells increased by 43.29% (P <0.01) in HuCCT1 cells treated with 5μmol / L PGE2 for 24 h. The expression of ICAM-1 protein in HuCCT1 cells treated with 10μmol / L EP1-4 agonist for 24 h was significantly higher than that of the control group (P <0.05). EP2 receptor agonist increased by 257.88% (P <0.05) L of EP2 receptor agonist for 24 h, the invasive ability of HuCCT1 cells was increased by 56.99% (P <0.01). The expression of ICAM-1 protein in HuCCT1 cells treated with 10 μmol / L EP2 receptor inhibitor AH6809 was significantly lower than that of PGE2 Group decreased by 49.14% (P <0.05), cell invasive ability decreased by 52.06% (P <0.01). The ICAM-1 protein expression decreased by 72.87% (P <0.05) and 80.78% (P <0.05) in the HuCCT1 cells treated with 10μmol / L AC inhibitor SQ22536 and 10μmol / L PKA inhibitor H89 <0.05). CONCLUSION: PGE2 can up-regulate the expression of ICAM-1 in HuCCT1 cells by activating the cAMP-PKA signal transduction pathway via EP2 receptor, thereby promoting the invasion and metastasis of HuCCT1 cells.