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目的:研究表皮生长因子受体(EGFR)抑制剂AG1478对子宫内膜癌细胞增殖和上皮-间质转化的影响。方法:蛋白印迹法检测子宫内膜癌细胞上皮标记蛋白E-cadherin、α-catenin,间质标记蛋白N-cadherin、vimentin及MMP-9、MMP-2蛋白的表达水平。MTT法检测不同浓度AG1478对子宫内膜癌细胞的增殖抑制作用。蛋白印迹法检测AG1478对子宫内膜癌细胞E-cadherin、α-catenin,N-cadherin、vimentin及MMP-9、MMP-2蛋白表达水平的影响。体外迁移实验检测AG1478对子宫内膜癌细胞迁移能力的影响。结果:(1)EGFR过表达可下调子宫内膜癌细胞上皮标记蛋白并上调间质标记蛋白及MMP-9、MMP-2蛋白的表达水平(P<0.05)。(2)AG1478对两种子宫内膜癌细胞均有增殖抑制作用,以经转染稳定过表达EGFR的Ishikawa细胞更为敏感(P<0.05)。(3)AG1478上调子宫内膜癌细胞上皮标记蛋白并下调间质标记蛋白及MMP-9、MMP-2蛋白的表达水平,以转染EGFR的Ishikawa细胞变化更为显著(P<0.05)。(4)AG1478可减弱子宫内膜癌细胞的迁移能力,以过表达EGFR的Ishikawa细胞更为明显(P<0.05)。结论:EGFR抑制剂AG1478可有效地抑制子宫内膜癌细胞增殖和上皮-间质转化。
Objective: To study the effect of epidermal growth factor receptor (EGFR) inhibitor AG1478 on proliferation and epithelial-mesenchymal transition of endometrial carcinoma cells. Methods: The expressions of E-cadherin, α-catenin, N-cadherin, vimentin, MMP-9 and MMP-2 in endometrial carcinoma cells were detected by Western blotting. MTT assay of different concentrations of AG1478 on the proliferation of endometrial cancer cells. The effect of AG1478 on protein expression of E-cadherin, α-catenin, N-cadherin, vimentin, MMP-9 and MMP-2 in endometrial carcinoma cells was detected by Western blotting. Effect of AG1478 on Migration of Endometrial Carcinoma Cells in Vitro by Migration Assay. Results: (1) EGFR overexpression downregulated the expression of epithelial marker protein and increased the expression of interstitial markers and MMP-9, MMP-2 protein in endometrial carcinoma cells (P <0.05). (2) AG1478 inhibited the proliferation of both endometrial cancer cells and was more sensitive to transfected Ishikawa cells stably overexpressing EGFR (P <0.05). (3) AG1478 up-regulated the expression of epithelial marker protein and down-regulated the expression of interstitial markers and MMP-9, MMP-2 protein in Ishikawa cells transfected with EGFR (P <0.05). (4) AG1478 attenuated the migration of endometrial cancer cells, and was more obvious in Ishikawa cells overexpressing EGFR (P <0.05). Conclusion: EGFR inhibitor AG1478 can effectively inhibit the proliferation of endometrial cancer cells and epithelial-mesenchymal transition.