Department of Oral and Maxillofacial Plastic Surgery, University Hospital Würzburg, Pleicherwall 2, 97070, Würzburg, Germany.
Institute of Pathology, University Würzburg, Josef-Schneider-Straße 2, 97080, Würzburg, Germany.
Clin Oral Investig. 2016 May;20(4):759-69. doi: 10.1007/s00784-015-1566-5. Epub 2015 Aug 23.
The objective of this study is to examine the efficacy of erlotinib and gefitinib with respect to epidermal growth factor (EGF) and cetuximab response in head and neck cancer cell lines.
Five human head and neck carcinoma cell lines were treated with EGF, cetuximab, erlotinib, and gefitinib, and the effects were measured with a crystal violet assay. The efficacies of cetuximab, erlotinib, and gefitinib in clinically relevant concentrations were statistically analyzed. The expression of the epidermal growth factor receptor (EGFR) and phosphorylation patterns were detected with fluorescence-activated cell sorting (FACS) analysis and western blot analysis. The endogenous production of EGF by the cells was detected with an enzyme-linked immunosorbent assay. Finally, EGFR, KRAS, BRAF, and PI3K mutation analyses were performed.
All of the cell lines had a poor or no response to EGF but exhibited distinct EGFR phosphorylation and EGFR expression. Compared to cetuximab, erlotinib and gefitinib demonstrated a greater impact on the majority of the cell lines. The only cell line that showed a concentration-dependent behavior toward EGF and strong EGFR phosphorylation was entirely resistant to cetuximab, erlotinib, and gefitinib. The production of EGF in all cell lines was very low. Mutational analysis of all cell lines revealed wild-type EGFR, KRAS, BRAF, and PI3K.
The prediction of anti-EGFR treatment cannot be based on responsiveness to EGF or EGFR activation.
Erlotinib and gefitinib show good response in EGF-independent cell lines and might be useful drugs in tumors that are less responsive to cetuximab.
本研究旨在探讨表皮生长因子(EGF)和西妥昔单抗对表皮生长因子受体(EGFR)的反应与厄洛替尼和吉非替尼疗效的关系。
用 EGF、西妥昔单抗、厄洛替尼和吉非替尼处理 5 个人类头颈部癌细胞系,并用结晶紫法测量其效果。对西妥昔单抗、厄洛替尼和吉非替尼在临床相关浓度下的疗效进行了统计学分析。用荧光激活细胞分选(FACS)分析和 Western blot 分析检测 EGFR 的表达和磷酸化模式。用酶联免疫吸附试验(ELISA)检测细胞内源性 EGF 的产生。最后,进行 EGFR、KRAS、BRAF 和 PI3K 突变分析。
所有细胞系对 EGF 反应较差或无反应,但表现出明显的 EGFR 磷酸化和 EGFR 表达。与西妥昔单抗相比,厄洛替尼和吉非替尼对大多数细胞系的影响更大。唯一对 EGF 表现出浓度依赖性和强 EGFR 磷酸化的细胞系对西妥昔单抗、厄洛替尼和吉非替尼完全耐药。所有细胞系的 EGF 产生量均非常低。对所有细胞系的突变分析均显示 EGFR、KRAS、BRAF 和 PI3K 均为野生型。
不能根据对 EGF 的反应性或 EGFR 激活来预测抗-EGFR 治疗。
厄洛替尼和吉非替尼在 EGF 非依赖性细胞系中显示出良好的反应,可能对西妥昔单抗反应性较低的肿瘤有用。