OBJECTIVE:To determine whether the combination of prostaglandin E2 (PGE2) and EP2, the subtype receptor of PGE2, could trans-activate the epidermal growth factor receptor (EGFR).
PATIENTS AND METHODS:In this experiment, we selected epithelial cells from normal esophageal mucosa as the negative control group, and the ESCC EC109 and TE-1 cell strain as the observation group. Real-time PCR and Western-blotting were used to detect the expression of EP2, EGFR and phosphorylated EGFR (p-EGFR). The pre-treatment of ESCC cell strains was carried out using Butaprost (special agonist of PGE2 and EP2) and RNAi of EP2, and we observed the expression of EP2, EGFR, and p-EGFR. WST-8 (CCK-8) was applied for the detection of the cell proliferation rate. The transwell invasion experiment was conducted for the detection of the invasion capability of cells. The expression of MMP-9 (matrix metalloproteinase-9), VEGF (vascular endothelial growth factor), pro-inflammatory factors (IL-6 and TNF-α) in the cell supernatant were detected using ELISA.
RESULTS:The high mRNA and protein expression of EP2, EGFR, and p-EGFR were found in the EC109 and TE-1 cell strains in the observation group, which were higher than those in the control group (p < 0.05). After the intervention of PGE2, EP2 expression was decreased and the p-EGFR expression was increased (p < 0.05). There was no variation found in the expression of EGFR (p > 0.05). After cells were intervened using Butaprost, the expressions of EP2 and p-EGFR were increased (p < 0.05), and there were no changes identified in the expression of EGFR (p > 0.05). After the intervention of RNAi, the expression of EP2 and p-EGFR was decreased (p < .05), and no changes were identified in the expression of EGFR (p > 0.05). After the intervention of PGE2 and Butaprost, great increases were seen in the cell proliferation rate, invasion capability, and the expression of MMP-9, VEGF, IL-6, and TNF-α in EC109 and TE-1 cell strains (p < 0.05), however, the intervention of RNAi could reduce above indexes (p < 0.05).
CONCLUSIONS:Through cell experiments, we verified that the combination of prostaglandin E2 (PGE2) and EP2, the subtype receptor of PGE2, could trans-activate the epidermal growth factor receptor (EGFR) to regulate the proliferation and invasion capability of esophageal squamous cell carcinoma (ESCC) cells, and secrete and express multiple cytokines, thus discovering the pathological mechanism of inflammation to carcinoma transition in the occurrence of ESCC, and providing the experimental evidence for the search of new target in the treatment of ESCC. ESCC cells can highly express the receptor subtype EP2 of PGE2 that can transactivate the EGFR, through which PGE2 is involved in the transition mechanism from inflammation to cancer.
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【Investigation on the regulatory effect of PGE2 on ESCC cells through the trans-activation of EGFR by EP2 and the relevant mechanism.].
【通过EP2对EGFR的反式激活，研究PGE2对ESCC细胞的调控作用及其机制。】
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DOI：10.26355/eurrev_201712_14011文章类型：杂志文章
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目的：确定前列腺素E2（PGE2）和EP2（PGE2的亚型受体）的组合是否可以反激活表皮生长因子受体（EGFR）。
病人与方法：本实验以正常食管粘膜上皮细胞为阴性对照组，以ESCC EC109和TE-1细胞株为观察组。实时PCR和Western-blotting用于检测EP2，EGFR和磷酸化EGFR（p-EGFR）的表达。使用Butaprost（PGE2和EP2的特殊激动剂）和EP2的RNAi进行ESCC细胞株的预处理，我们观察到EP2，EGFR和p-EGFR的表达。 WST-8（CCK-8）用于检测细胞增殖率。进行transwell侵袭实验以检测细胞的侵袭能力。使用ELISA检测细胞上清液中MMP-9（基质金属蛋白酶-9），VEGF（血管内皮生长因子），促炎因子（IL-6和TNF-α）的表达。
结果：观察组EC109和TE-1细胞株中EP2，EGFR和p-EGFR的mRNA和蛋白表达较高，高于对照组（p <0.05）。在PGE2干预后，EP2表达降低，而p-EGFR表达升高（p <0.05）。在EGFR的表达中没有发现变化（p＞ 0.05）。用Butaprost干预细胞后，EP2和p-EGFR的表达增加（p <0.05），而EGFR的表达没有变化（p> 0.05）。 RNAi干预后，EP2和p-EGFR的表达降低（p <.05），而EGFR的表达未见变化（p> 0.05）。在PGE2和Butaprost的干预下，EC109和TE-1细胞株的细胞增殖速率，侵袭能力以及MMP-9，VEGF，IL-6和TNF-α的表达均显着增加（p < 0.05），但是，RNAi的干预可以降低上述指标（p <0.05）。
结论：通过细胞实验，我们证实前列腺素E2（PGE2）和EP2（PGE2的亚型受体）的组合可以反激活表皮生长因子受体（EGFR）来调节食管鳞状细胞癌的增殖和侵袭能力。 （ESCC）细胞，并分泌和表达多种细胞因子，从而发现了发生ESCC时炎症向癌变的病理机制，为寻找ESCC的新靶标提供了实验依据。 ESCC细胞可以高表达PGE2的受体亚型EP2，该亚型可以使EGFR活化，PGE2可以通过该亚型参与从炎症到癌症的转变机制。
病人与方法：本实验以正常食管粘膜上皮细胞为阴性对照组，以ESCC EC109和TE-1细胞株为观察组。实时PCR和Western-blotting用于检测EP2，EGFR和磷酸化EGFR（p-EGFR）的表达。使用Butaprost（PGE2和EP2的特殊激动剂）和EP2的RNAi进行ESCC细胞株的预处理，我们观察到EP2，EGFR和p-EGFR的表达。 WST-8（CCK-8）用于检测细胞增殖率。进行transwell侵袭实验以检测细胞的侵袭能力。使用ELISA检测细胞上清液中MMP-9（基质金属蛋白酶-9），VEGF（血管内皮生长因子），促炎因子（IL-6和TNF-α）的表达。
结果：观察组EC109和TE-1细胞株中EP2，EGFR和p-EGFR的mRNA和蛋白表达较高，高于对照组（p <0.05）。在PGE2干预后，EP2表达降低，而p-EGFR表达升高（p <0.05）。在EGFR的表达中没有发现变化（p＞ 0.05）。用Butaprost干预细胞后，EP2和p-EGFR的表达增加（p <0.05），而EGFR的表达没有变化（p> 0.05）。 RNAi干预后，EP2和p-EGFR的表达降低（p <.05），而EGFR的表达未见变化（p> 0.05）。在PGE2和Butaprost的干预下，EC109和TE-1细胞株的细胞增殖速率，侵袭能力以及MMP-9，VEGF，IL-6和TNF-α的表达均显着增加（p < 0.05），但是，RNAi的干预可以降低上述指标（p <0.05）。
结论：通过细胞实验，我们证实前列腺素E2（PGE2）和EP2（PGE2的亚型受体）的组合可以反激活表皮生长因子受体（EGFR）来调节食管鳞状细胞癌的增殖和侵袭能力。 （ESCC）细胞，并分泌和表达多种细胞因子，从而发现了发生ESCC时炎症向癌变的病理机制，为寻找ESCC的新靶标提供了实验依据。 ESCC细胞可以高表达PGE2的受体亚型EP2，该亚型可以使EGFR活化，PGE2可以通过该亚型参与从炎症到癌症的转变机制。
