摘要
目的:探讨表皮生长因子受体(EGFR)的单克隆抗体西妥昔单抗(Cetuximab,C225)与化疗药物联合作用于大肠癌和肺癌细胞的最佳联合方案及其联合机制。方法:采用MTT法测定不同药物单药、两药及三药联合分别对大肠癌细胞LoVo和肺腺癌细胞A549的IC50值;计算不同联合方案的联合指数(combination index,CI),以确定最佳联合方案;Western blot法检测不同给药方案对EGFR下游信号通路的关键分子AKT、MAPK磷酸化表达的影响;流式细胞仪检测不同联合方案对细胞周期分布及凋亡比例的影响。结果:C225与化疗药物联合可增强其对细胞的杀伤作用,且以先化疗药物、后C225的序贯方案协同作用最强。先化疗药物、后C225的序贯方案对AKT、MAPK磷酸化的抑制明显强于其它联合方案,并且能诱导更多肿瘤细胞发生凋亡以及产生G2/M期阻滞。结论:C225与化疗药物联合存在最佳联合方案,其机制可能与药物对EGFR下游信号通路关键分子的磷酸化抑制及促进细胞凋亡和影响细胞周期分布有关。
Objective：To determine the best combination schedule and related mechanisms of cetuximab and cytotoxic agents. Methods：The colon cancer cell line I.oVo and lung cancer cell line A549 was used. Inhibitions of cell proliferation were evaluated by MTT assay. The combination effect was determined by combination index analysis. The phosphorylation expression of MAPK and AKT was evaluated by western blot analysis. The cell cycle and apoptosis was analyzed by FACS. Results： The combination effect of the cetuximab and cytotoxic agents was sequence-dependent. The synergistic effect was obtained when cytotoxic agents were given before cetuximab. When these two agents were given at the same time, the combination effect turned to be antagonism. The synergistic effect was related to completely inhibition of the phosphorylation expression of MAPK and AKT. Moreover, cell cycle arrest at G2/M phase and more apoptosis was induced by synergistic combination. Conclusion：These findings suggest the synergistic combination of cetuximab and cytotoxic agents was sequence depended.
出处
《临床肿瘤学杂志》
CAS
2007年第10期736-739,共4页
Chinese Clinical Oncology
基金
全军"十一.五"科技攻关课题(06G140)
