双靶点阻滞对肺腺癌细胞的协同抑制作用及其可能机制  

Synergistic inhibitory effect of blocking both EGFR-and COX-2-related pathways on lung cancer A549 cells and its underlying mechanism

作　　者：王亚丽[1] 吴峰[1] 牟晓燕[2] 胡锋[1] 陶玉坚[1] 黄玉民[1] 胡涛[1] 邵向荣[1] 

机构地区：[1] 扬州市第一人民医院呼吸科,扬州225002 [2] 山东大学附属省立医院呼吸科,济南250021

出　　处：《中华老年多器官疾病杂志》2016年第12期932-937,共6页Chinese Journal of Multiple Organ Diseases in the Elderly

基　　金：山东省自然科学基金(ZR2009CM1250)~~

摘　　要：目的：探讨联合阻断表皮生长因子受体（EGFR）和环氧合酶－2（COX－2）对肺腺癌A549细胞株的协同抑制作用及其可能机制。方法采用不同药物干预肺癌A549细胞株，分为4组：正常对照组、单药吉非替尼组、单药塞来昔布组、联合用药组。药物干预细胞48 h后台盼蓝（trypan blue）染色法检测药物对细胞生长的影响；以Hoechst33258染色法和流式细胞术观察用药前后细胞凋亡和细胞周期的变化。采用Western blot检测EGFR和COX－2蛋白的表达情况。结果随着时间和剂量增加，单药吉非替尼与塞来昔布对A549细胞的抑制作用增强，加药48 h，联合用药组抑制率明显高于单药组（P＜0．01）。联合用药组细胞凋亡率明显高于单独用药组（32．40％vs 7．12％和8．43％；P＜0．01）。联合用药组S期细胞比例为（3．2±0．9）％，较单药吉非替尼组[（37．4±1．6）％]和单药塞来昔布组[（21．0±3．1）％]明显减少（P＜0．01）；联合用药组G0／G1期细胞比例为（87．2±6．4）％，较单药吉非替尼组[（61．4±5．2）％]和单药塞来昔布组[（51．8±4．7）％]明显增加（P＜0．01）。与单独用药组比较，联合用药组EGFR和COX－2蛋白的表达明显减弱（P＜0．05）。结论联合用药通过EGFR和COX－2双靶点阻滞发挥作用，有望为肺癌的化学预防和治疗提供新的策略。Objective To determine the synergic effect of simultaneously blocking epidermal growth factor receptor （ EGFR） and cyclooxygenase-2 on the growth inhibition of human lung cancer A 549 cells.Methods The A549 cells were treated by gefitinib （5 μmol/L）, celecoxib （25 μmol/L）, or a combination of them at the same dosages respectively for 48 h, and the cells without treat-ment served as control .Cell growth was detected by trypan-blue exclusion assay , apoptosis rate and cell cycle were measured by flow cytometry and Hoechst33258 staining, expression of EGFR and COX-2 proteins was determined by Western blotting .Results With time and dose increase , both gefitinib and celecoxib showed stronger inhibitory effect on the growth of A 549 cells, and the combination of them had more significant inhibitory effect than the gefitinib and celecoxib groups （P〈0.01）.The combination group also had obvious higher apoptosis rate than the monotherapy groups （32.40%vs 7.12%and 8.43%, P〈0.01）.The proportion of the cells at S stage was （3.2 ±0.9）%in the combination group , significantly decreased when compared with those in the gefitinib and celecoxib groups [（37.4 ±1.6）%, （21.0 ±3.1）%, P〈0.01].The combination group had obviously more cells arrested at G0/G1 stage than the monotherapy groups [（87.2 ±6.4）% vs （61.4 ±5.2）% and （51.8 ±4.7）%, P〈0.01].The protein levels of EGFR and COX-2 were significantly lower in the combination group than the other 2 groups （P〈0.05）.Conclusion The combination treatment of gefitinib and celecoxib exerts inhibitory effect through blocking both EGFR-and COX-2-related pathways , and the regimen might provide a promising strategy for cancer therapy and chemoprevention in lung cancer .

关 键 词：塞来昔布 吉非替尼 细胞周期 细胞凋亡 表皮生长因子受体 环氧合酶-2 

分 类 号：R734.2[医药卫生—肿瘤]