Unit of Cell and Molecular Biology, Dundee Dental School, University of Dundee, Dundee, UK.
J Oral Pathol Med. 2021 Nov;50(10):1031-1039. doi: 10.1111/jop.13230. Epub 2021 Aug 17.
We investigated the role of epidermal growth factor (EGF) and transforming growth factor α (TGFα) on Asian head and neck cancer patient cell lines; in terms of epithelial-to-mesenchymal transition (EMT) and cell migration to determine whether these changes could be reversed using tyrosine kinase inhibitors (Gefitinib and Erlotinib).
Cell migration, protrusion and EMT were assessed using both Scatter assay and Scratch assay. Protein expression and localisation were evaluated using immunofluorescence, SDS-PAGE and Western blotting techniques to identify the involvement of phosphorylated MAPK (Thr202/Tyr204), phosphorylated EGFR (Y1068) and phosphorylated AKT (Ser473) protein expression.
EGF and TGFα induced an EMT-like phenotypical change, cellular protrusion and cell migration while Gefitinib and Erlotinib blocked these morphological changes and cell migration. We also examined the effect of EGF/TGF α± tyrosine kinase inhibitors on phosphorylation sites Y1068 of epidermal growth factor receptor (EGFR). Y1068 was phosphorylated in all test conditions, and all tested concentrations of inhibitors did not inhibit Y1068 phosphorylation. EGF and TGFα increased phosphorylation of MAPK (Thr202/Tyr204) residues compared with serum-free control while a one-hour pre-treatment with tyrosine kinase inhibitor(s) before addition of growth factors completely blocked this phosphorylation. Phosphorylation of Akt Ser 473 was also induced by EGF and TGFα, and a one-hour pre-treatment with the tyrosine kinas inhibitor(s) reduced this phosphorylation.
These data suggest that Gefitinib and Erlotinib prevent activation of downstream signalling proteins MAPK (Thr202/Tyr204) and Akt (Ser473) thereby blocking phenotypic change and cell migration. This study supports the potential therapeutic value of Gefitinib and Erlotinib in targeting head and neck cancer.
我们研究了表皮生长因子 (EGF) 和转化生长因子 α (TGFα) 在亚洲头颈部癌症患者细胞系中的作用；研究了上皮间质转化 (EMT) 和细胞迁移，以确定这些变化是否可以使用酪氨酸激酶抑制剂 (吉非替尼和厄洛替尼) 逆转。
使用散射分析和划痕分析评估细胞迁移、突起和 EMT。使用免疫荧光、SDS-PAGE 和 Western blot 技术评估蛋白表达和定位，以确定磷酸化 MAPK（Thr202/Tyr204）、磷酸化 EGFR（Y1068）和磷酸化 AKT（Ser473）蛋白表达的参与情况。
EGF 和 TGFα 诱导 EMT 样表型变化、细胞突起和细胞迁移，而吉非替尼和厄洛替尼阻断这些形态变化和细胞迁移。我们还研究了 EGF/TGFα±酪氨酸激酶抑制剂对表皮生长因子受体 (EGFR) 的 Y1068 磷酸化位点的影响。在所有测试条件下，Y1068 均发生磷酸化，并且所有测试浓度的抑制剂均未抑制 Y1068 磷酸化。与无血清对照相比，EGF 和 TGFα 增加了 MAPK（Thr202/Tyr204）残基的磷酸化，而在添加生长因子之前用酪氨酸激酶抑制剂预处理一小时完全阻断了这种磷酸化。EGF 和 TGFα 还诱导 Akt Ser 473 的磷酸化，用酪氨酸激酶抑制剂预处理一小时可降低这种磷酸化。
这些数据表明，吉非替尼和厄洛替尼可阻止下游信号蛋白 MAPK（Thr202/Tyr204）和 Akt（Ser473）的激活，从而阻断表型变化和细胞迁移。这项研究支持吉非替尼和厄洛替尼在头颈部癌症治疗中的潜在治疗价值。
