厄洛替尼和培美曲塞不同方式联合对肺腺癌A549细胞的作用  

Effect of different combination protocols of pemetrexed and erlotinib on the lung adenocarcinoma A549 cells

作　　者：孙杰[1] 牟晓燕[1] 董雪丽[1] 

机构地区：[1]山东大学附属省立医院保健呼吸科,山东济南250014

出　　处：《肿瘤》2013年第12期1039-1046,共8页Tumor

基　　金：山东省科技发展计划项目(编号:2010GSF10253);山东省自然科学基金资助项目(编号:ZR2009CM125)

摘　　要：目的 :探讨培美曲塞与厄洛替尼联合及序贯应用对肺腺癌A549细胞增殖和凋亡的影响及其可能的机制。方法 :培美曲塞和厄洛替尼单药、联合及序贯作用A549细胞72 h后,MTT法检测细胞的增殖并计算联合指数,FCM法检测细胞周期分布和细胞凋亡率,蛋白质印迹法检测磷酸化细胞外调节蛋白激酶1/2(phospho-extracellular regulated protein kinase 1/2,p-ERK1/2)、磷酸化-AKT(phospho-AKT,p-AKT)和磷酸化表皮生长因子受体(phospho-epidermal growth factor receptor,p-EGFR)的表达。结果 :培美曲塞序贯厄洛替尼对抑制A549细胞的增殖具有协同作用,厄洛替尼联合或序贯培美曲塞对A549细胞的增殖具有拮抗作用。与对照组(未进行药物干预的A549细胞)相比,培美曲塞组和培美曲塞序贯厄洛替尼组的S期细胞所占比例增多(P<0.05),厄洛替尼组、厄洛替尼联合培美曲塞组及厄洛替尼序贯培美曲塞组的G1期细胞所占比例增多(P<0.05)。与厄洛替尼联合培美曲塞组及厄洛替尼序贯培美曲塞组相比,培美曲塞序贯厄洛替尼组A549细胞的凋亡率明显增加(P<0.05)。与对照组比较,培美曲塞序贯厄洛替尼组A549细胞p-AKT和p-EGFR的表达水平明显下调(P<0.05),而p-ERK1/2的表达水平差异无统计学意义(P>0.05)。结论 :培美曲塞序贯厄洛替尼对A549细胞的增殖具有协同作用,其机制可能与磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase,PI3K)-AKT通路的表达有关。Objective： To investigate the effect of combination and sequensial application of pemetrexed and erlotinib on the proliferation and apoptosis of lung adenocarcinoma A549 cells and to explore its possible mechanism. Methods： After A549 cells were treated with pemetrexed or erlotinib alone, or in combination or sequensial application for 72 h, the cell proliferation rate was measured by MTT assay and the combination index was calculated. Cell cycle distribution and apoptosis rate were detected by flow cytometry. The expressions of phospho-extracellular regulated protein kinase 1/2 （p-ERK1/2）, phospho- AKT （p-AKT） and phospho- epidermal growth factor receptor （p-EGFR） were detected by Western blotting. Results： The application of erlotinib following pemetrexed had a synergistic effect on A549 cell proliferation while pemetrexed following or combined with erlotinib had an antagonistic effect. As compared with the control group （A549 cells without treatment）, the percentage of A549 cells in S phase after treatment with pemetrexed or erlotinib following pemetrexed was increased （P 〈 0.05）, and the percentage of A549 cells in G1 phase after treatment with erlotinib or pemetrexed following or combined with erlotinib was increased （P 〈 0.05）. As compared with pemetrexed following erlotinib group or pemetrexed in combination with erlotinib group, the apoptosis rate of A549 cells after treatment with erlotinib following pemetrexed was increased （P 〈 0.05）. As compared with the control group, the expression levels of p-EGFR and p-AKT in A549 cells after treatment with erlotinib following pemetrexed were down-regulated （P 〈 0.05）, and the expression level of p-ERK1/2 had no significant difference （P 〉 0.05）. Conclusion： The application of erlotinib following pemetrexed has a synergisticeffect on A549 cell proliferation. This effect may be related to the expression of phosphatidylinositol 3-kinase （PI3K）-AKT signal pathway.

关 键 词：肺肿瘤 药物疗法 联合 体外研究 厄洛替尼 培美曲塞 

分 类 号：R734.2[医药卫生—肿瘤]