舒尼替尼联合多西他赛序贯给药作用于EGFR-TKIs抵抗的非小细胞肺癌A549细胞株的效应  

Schedule-dependent cytotoxic synergism of docetaxel and sunitinib in EGFR-TKIs-resistant human non-small cell lung cancer cell line A549

作　　者：潘峰[1] 田静[2] 张旭超 潘跃银[1] 

机构地区：[1]安徽医科大学第一附属医院肿瘤科,合肥230022 [2]安徽医科大学公共卫生学院流行病与卫生统计学系,合肥230032 [3]广东省肺癌研究所中心实验室,广州510080

出　　处：《肿瘤》2011年第9期806-812,共7页Tumor

基　　金：安徽省科技计划重点项目(编号:07020304102)

摘　　要：目的:本研究旨在探索舒尼替尼单药对表皮生长因子受体酪氨酸激酶抑制剂(epidermal growth factor receptor-tyrosine kinase inhibitors,EGFR-TKIs)耐药的非小细胞肺癌A549细胞的效应,以及联合多西他赛不同给药顺序对A549细胞株的不同效应,揭示产生这种差异的机制。方法:MTT法检测舒尼替尼和多西他赛对A549细胞的半数抑制浓度(half maximal inhibitory concentration,IC50),以及不同给药顺序的联合指数(combination index,CI)的变化。FCM检测舒尼替尼(3.5μmol/L)和多西他赛(5.0nmol/L)不同联合方式作用后的A549细胞周期变化。蛋白质印迹法检测不同给药顺序作用后细胞中p-AKT蛋白水平的变化。结果:舒尼替尼和多西他赛作用于A549细胞72h的IC50分别为3.6±0.41μmol/L和5.5±0.95nmol/L;且两者的联合抗增殖作用与给药顺序有关,先用多西他赛后再用舒尼替尼表现出明显的协同效应,而先用舒尼替尼后再用多西他赛和同时给药则产生拮抗作用。FCM检测结果显示,舒尼替尼和多西他赛分别将A549细胞阻滞于G0/G1期和S期;当先用多西他赛后再用舒尼替尼时,S期细胞显著增多,G0/G1期细胞则显著减少。蛋白质印迹结果显示,先用多西他赛后再用舒尼替尼时p-AKT/AKT比值下降最多。结论:舒尼替尼能抑制A549细胞的生长,先用多西他赛后再用舒尼替尼能产生明显的协同效应;推测产生这种差异的机制可能与细胞周期和生存通路蛋白表达的变化相关。Objective：To explore the efficacy of sunitinib as a single agent and in combination with docetaxel under three different sequence-dependent schedules on epidermal growth factor receptor tyrosine kinase inhibitors（EGFR-TKIs）-resistant human non-small cell lung cancer A549 cells,and to reveal the responsible mechanisms for generating these differences.Methods：The half inhibitory concentration（IC50） and the combination index（CI） values of docetaxel or sunitinib alone as well as in combination under three different sequence-schedules for A549 cells were estimated by MTT.The flow cytometry（FCM） was used to analyze the alteration of cell cycle in A549 cells treated with sunitinib（3.5 μmol/L） and docetaxel（5.0 nmol/L） by different administration programs.The change of p-AKT protein level under different sequence-schedules was measured by Western blotting.Results：At 72 h,the IC50 values of docetaxel and sunitinib for A549 cells were 3.6±0.41 μmol/L and 5.5±0.95 nmol/L,respectively.The synergic effect of combination therapy with docetaxel and sunitinib was associated with the sequence of administration.The sequence of docetaxel followed by sunitinib showed a significant synergic effect;however,when the two drugs were used concurrently or by the reverse sequence,they illustrated an antagonistic effect.FCM analysis showed that sunitinib resulted in cell cycle arrest at G0/G1 phase,while docetaxel induced cell cycle arrest at S phase.When docetaxel was administered followed by sunitinib,the percentages of S-phase and G0/G1-phase cells were significantly increased and decreased,respectively.The ratio of p-AKT/AKT reached its lowest level in A549 cells treated with docetaxel followed by sunitinib.Conclusion：Sunitinib can inhibit the growth of A549 cells.The sequence of docetaxel followed by sunitinib can produce significant synergism.The mechanism underlying the difference may be related to the changes of cell cycle and the survival pathway.

关 键 词：癌 非小细胞肺 药物疗法 联合 用药计划表 细胞周期 蛋白质丝氨酸苏氨酸激酶 舒尼替尼 多西他赛 

分 类 号：R734.2[医药卫生—肿瘤]