【摘要】 背景与目的:表皮生长因子受体(epidermal growth factor receptor,EGFR)在非小细胞肺癌中高度表达并且参与其病理过程。目前针对于表皮生长因子受体开发了两种治疗肺癌的药物,一种是EGFR小分子抑制剂吉非替尼(Gefitinib),另一种是EGFR单克隆抗体西妥昔单抗(Cetuximab)。然而,Gefitinib、Cetuximab联合治疗肺癌的效果以及具体机制还不是很清楚。本实验旨在研究Gefitinib、Cetuximab对人肺癌细胞A549、H460、H1299、SPC-A、95C、95D细胞增殖、细胞周期与凋亡、侵袭转移及相关蛋白表达的影响,并探讨其相应的分子机制。方法:CCK8法检测各细胞对Gefitinib、Cetuximab的IC50。流式细胞技术观察细胞凋亡状况;Transwell实验检测细胞侵袭和迁移黏附情况;Westernblot检测Gefitinib、Cetuximab对增殖相关信号通路蛋白表达的变化。结果:1nmol/LGefitinib和5μmol/LCetuximab对A549细胞生长抑制率大概为30%和40%,而两者联合使用的抑制率能够达到50%以上。5μmol/LGefitinib和10nmol/LCetuximab单独及联合作用对A549细胞迁移率降低约为18%、22%和35%。通过Westernblot检测到Gefitinib、Cetuximab处理组的p-AKT、p-EGFR、p-MAPK蛋白量较对照组明显下降。结论:Gefitinib和Cetuximab能够抑制肺癌细胞生长和迁移,并且有着良好的协同作用,提示两药联合使用临床肺癌治疗可能具有很大的潜力。更多还原

【Abstract】 Background and purpose:Epidermal growth factor receptor(EGFR) is frequently overexpressed in non-small cell lung cancer(NSCLC) and has been implicated in the pathogenesis of the disease.Presently, a small-molecule inhibitor(Gefitinib) and a monoclonal antibody of EGFR(Cetuximab) have been developed as the 2 main drugs for therapeutic targeting of EGFR.However, the therapeutic effect and mechanism of Gefitinib and Cetuximab still need to be investigated.In our research, we aimed to investigate the apoptosis, invasion, and metastasis of A549, H460, H1299, SPC-A, 95C, 95D cells in the presence of Gefitinib, Cetuximab treatment.Also, their molecular mechanisms would all be explored.Methods:IC50 of A549, H460, H1299, SPC-A, 95C, 95D cells that response to Gefitinib and Cetuximab would be studied by CCK8 assay.The invasion and metastasis of different lung cancer cells were investigated through the Transwell method.Western blot was applied to study the expression of EGFR signal pathway related proteins.Results:1 nmol/L Gefitinib or 5 μmol/L Cetuximab could each inhibit the growth of A549 by about 30% to 40%, but when combined, the inhibition rate increased to 50% up.5 μmol/L Gefitinib, 10 nmol/ L Cetuximab, or a combination of both, decreased the invasion rate by about 18%, 22%, and 35%, respectively.The expression of p-AKT, p-EGFR, and p-MAPK were reduced significantly through Gefitinib and Cetuximab treatments with Western blot.Conclusion:Gefitinib and Cetuximab could inhibit the growth and invasion of lung cancer cells significantly and with synergistic effects.A combination of Gefitinib and Cetuximab is a potential strategy for lung cancer therapy.更多还原