摘要
目的探讨蛋白酶体抑制剂MG132联合顺铂对食管鳞癌细胞的杀伤作用及其机制。方法分别以顺铂（100mg／L）、MG132（5μmol／L）及顺铂＋MG132处理食管鳞癌EC9706细胞24h，细胞计数试剂盒（CCK-8）法检测细胞活力，膜联蛋白V（AnnexinV）-异硫氰酸荧光素（FITC）细胞凋亡检测试剂盒定量检测细胞凋亡率，Western blot检测凋亡相关蛋白半胱氨酰天冬氨酸特异性蛋白酶（Caspase）-8、Caspase-3和核转录因子（NF）-κB的表达。结果MG132可以抑制肿瘤细胞的增殖，增强顺铂对食管鳞癌细胞的杀伤作用，使细胞存活率从（68．19±3．41）％降低至（29．37±4．16）％（P〈0．05）。MG132联合顺铂处理组的细胞凋亡率为（68．45±2．58）％，明显高于单独处理组的（23．55±1．23）％和（25．87±2．07）％（P〈0．01）。联合用药组细胞Caspase-8、Caspase-3的表达水平高于顺铂处理组，但NF—κB较单独顺铂处理组明显下降（P〈0．05）。结论MG132通过抑制肿瘤细胞增殖，促进凋亡，提高顺铂对食管鳞癌细胞的杀伤作用。
Objective To investigate the antitumor activity of proteasome inhibitor MG132 in human esophageal squamous cancer cells. Methods EC9706 cells were treated with 100 mg/L cisplatin, 5 μmol/L MG132 and cisplatin ＋ MG132 for 24 h. Cell viability was measured by using cell counting kit-8 （ CCK-8 ） assay. The percentages of apoptotic cells were analyzed by using Annexin V-fluoresceine isothiocyanate （FITC） apoptosis detection kit. Western blotting assay was used to detect the expression of nuclear factor-κB （NF-KB） , Caspase-8 and -3. Results Exposure of cells to cisplatin combined with MG132 resuited in a marked increase in the cell cytotoxicity of EC9706 cells as compared with the single agent. The combined cisplatin and MG132 treatment induced more apoptosis in tumor cells than in cisplatin treatment alone （68. 45 ± 2. 58 vs. 23.5 ± 1.23% ;P 〈 0. 01 ）. MG132 significantly enhanced cisplatin-induced apoptosis in association with the activation of Caspase-3 and-8. These events were accompanied by the downregulation of NF-κB. Conclusion These findings demonstrate a novel mechanism by which proteasome inhibitor MG132 potentiates cisplatin-induced apoptosis in human esophageal squamous cell carcinoma.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2014年第5期1042-1044,共3页
Chinese Journal of Experimental Surgery
基金
河南省高校科教创新团队项目（13IRTSTHN011）
关键词
蛋白酶体抑制剂
顺铂
脱噬作用
食管鳞癌
Proteosome inhibitor
Cisplatin
Apoptosis
Esophageal squamous cell carcinoma
