口虾蛄提取物对人肺腺癌耐药细胞株A549/DDP增殖、耐药的影响及机制  

Effects of ESO extracts on proliferation and drug resistance of human lung adenocarcinoma drug-resistant cell line A549/DDP

作　　者：邵松军[1,2] 方海燕 段玲弟 袁国航 张湘燕[1] 郭兵[2] 

机构地区：[1]贵州医科大学附属省人民医院,贵阳550002 [2]贵州医科大学 [3]贵州省第二人民医院

出　　处：《山东医药》2017年第22期1-4,共4页Shandong Medical Journal

基　　金：贵州省科技合作计划项目(黔科合LH字2015-7126号)

摘　　要：目的观察口虾蛄乙酸乙酯提取物(ESO)对人肺腺癌耐药细胞株A549/DDP增殖、耐药的影响,并探讨其机制。方法选取人肺腺癌耐药细胞株A549/DDP,用不同浓度(0、200、400、800μg/m L)的ESO干预细胞,24 h后,采用MTT法测算细胞增殖抑制率,流式细胞技术测算细胞周期百分率。采用流式细胞技术观察400μg/m L ESO干预0、30、60 min细胞内荧光染料罗丹明123(Rh123)蓄积和外排情况。采用Western blotting法检测不同浓度ESO干预24 h后细胞多药耐药蛋白1(MDR1),并检测400μg/m L ESO干预2、4、8 h后细胞ERK1/2、p-ERK1/2蛋白。结果 200、400、800μg/m L的ESO干预细胞增殖抑制率分别为19.940%±0.041%、37.360%±0.024%、42.510%±0.008%,两两相比P均<0.01。0、200、400、800μg/m L ESO干预细胞G2/M期分别为24.06%±2.22%、31.10%±0.79%、35.64%±1.15%、43.19%±0.43%,两两相比P均<0.01。Rh123在ESO干预细胞内积累增多、外排减少,且60 min时较30 min时明显(P均<0.01)。0、200、400、800μg/m L ESO干预细胞MDR1蛋白相对表达量分别为1.22±0.23、0.83±0.16、0.37±0.09、0.31±0.04,两两相比P均<0.01。ESO干预不同时间A549/DDP细胞ERK1/2蛋白表达无变化,p-ERK1/2蛋白表达上调(P均<0.01)。结论 ESO通过使A549/DDP细胞发生周期阻滞,从而抑制其增殖;通过调节MAPK-ERK1/2信号通路,使其多药耐药发生逆转。Objective To observe the effects of ethyl acetate extracts of Squilla oratoria（ ESO） on proliferation and drug resistance of human lung adenocarcinoma drug-resistant cell line A549/DDP and to investigate its mechanism. Methods We selected the A549/DDP cell line,which were treated with different concentrations of ESO（ 0,200,400,800μg/ml） for 24 h. The proliferation inhibition rates of A549/DDP cells were detected by MTT assay. The cell cycle percentage was determined by flow cytometry（ FCM）. We used FCM to detect the Rhodamine123（ Rh123） accumulation and efflux in A549/DDP cells treated with 400 μg/m L ESO for 0,30,and 60 min. After the cells were treated with different concentrations of ESO for 24 h,the multidrug resistance 1（ MDR1） protein was detected by Western blotting. Meanwhile,we detected the ERK1/2,p-ERK1/2 protein of cells after 400 μg/m L ESO treatment for 2,4,and 8 h. Results The proliferation inhibition rates of A549/DDP cells treated with 200,400,and 800 μg/m L ESO were 19. 940% ± 0. 041%,37. 360% ± 0. 024%,and 42. 510% ± 0. 008%,and significant difference was found between every two groups（ all P〈0. 01）. A549/DDP cells treated with 0,200,400,and 800 μg/m L ESO could be blocked in G2/M phase,and the proliferation inhibition rates were 24. 06% ± 2. 22%,31. 1% ± 0. 79%,35. 64% ± 1. 15%,and 43. 19% ± 0. 43%,respectively; significant difference was found between every two groups（ all P〈0. 01）. ESO increased the accumulation of Rh123 level of A549/DDP cells,reduced the efflux,and this change was more significant at 60 min than at 30 min（ all P〈0. 01）. The expression of MR1 protein was 1. 22 ± 0. 23,0. 83 ± 0. 16,0. 37 ± 0. 09,and 0. 31 ± 0. 04 in A549/DDP cells after 0,200,400,800 μg/m L of ESO treatment respectively,all P〈0. 01. However,p-ERK1/2 protein expression was up-regulated in A549/DDP cells（ P〈0. 01）,but p-ERK1/2 protein expression had no variation at different time of ESO treatment. Conclusion ESO inhibits the proliferation of A549/D

关 键 词：口虾蛄 非小细胞肺癌 多药耐药1 细胞外调节蛋白激酶1/2蛋白 

分 类 号：R285.5[医药卫生—中药学]