盐霉素对人鼻咽癌CNE2/DDP细胞顺铂耐药的影响及其机制  

Effect of salinomycin on cisplatin resistance of human nasopharyngeal carcinoma cells CNE2/DDP and its mechanisms

作　　者：陈冬杰[1] 廖雪阳[1] 陈可绪[1] 李毅斌[1] 汪森明[1] 

机构地区：[1]南方医科大学珠江医院,广州510282

出　　处：《山东医药》2016年第48期20-23,共4页Shandong Medical Journal

基　　金：广州市科技计划项目(2011Y2-00019-3)

摘　　要：目的探讨盐霉素对人鼻咽癌CNE2/DDP细胞顺铂耐药的影响及其可能的作用机制。方法将人鼻咽癌CNE2/DDP细胞随机分为四组,分别加入2μg/m L顺铂(顺铂组)、8μmol/L盐霉素(盐霉素组)、2μg/m L顺铂和8μmol/L盐霉素(联合组)、0.1%二甲基亚砜(对照组)进行处理。采用CCK-8法检测各组细胞增殖抑制率,流式细胞术检测细胞凋亡率,Western blotting法检测Wnt/β-catenin信号通路(β-catenin、p-LRP6)、m TORc1信号通路(P70S6K、p-P70S6K、S6、p-S6)相关蛋白相对表达量,荧光分光光度法检测caspase3、caspase9活性。结果与对照组比较,顺铂组、盐霉素组、联合组细胞增殖抑制率、凋亡率和caspase3、caspase9活性均升高(P均<0.05);顺铂组、盐霉素组和联合组上述指标均逐渐升高,两两比较P均<0.05。β-catenin、p-LRP6、p-P70S6K、p-S6蛋白相对表达量:顺铂组和对照组均高于盐霉素组和联合组(P均<0.05),顺铂组与对照组、盐霉素组与联合组比较差异均无统计学意义(P均>0.05)。P70S6K、S6蛋白相对表达量:各组比较差异均无统计学意义(P均>0.05)。结论盐霉素可以逆转人鼻咽癌CNE2/DDP细胞的顺铂耐药;激活caspase活性、抑制Wnt/β-catenin和m TORc1信号通路可能是其作用机制。Objective To investigate the effect of salinomycin on the cisplatin resistance of human nasopharyngeal carcinoma CNE2/DDP cells and its possible mechanisms. Methods CNE2/DDP cells were randomly divided into four groups, which were separately treated with 2 μg/mL cisplatin （cisplatin group）, 8 μmmol/L salinomycin （salinomycin group）, 8μmmol/L salinomycin ＋ 2 μg/mL cisplatin （ combination group） and 0.1% DMSO （ control group）. Inhibitory rate of proliferation was assessed by the Cell Counting Kit 8. Apoptosis rate was measured by flow cytometry. Caspases3 and caspase9 activity was assessed by a colorimetric assay utilizing specific substrates. The protein expression of Wnt/β- catenin signaling pathway （β-catenin and p-LRP6） as well as mTORcl signaling pathway （P70S6K, p-P70S6K, $6, and p-S6） was evaluated by Western blotting. Results Compared with the control group, the inhibitory rate, apoptosis rate, Caspase-3/9 activity were increased in the cisplatin group, salinomycin group, and combination group （ all P 〈0.05）. In- dexes mentioned above were increased the most in the combination group while salinomycin group ranked the second and cisplatin group ranked the third （all P 〈0.05）. The expression of β-catenin, p-LRP6, p-P70S6K, and p-S6： the salino- mycin group and combination group was decreased as compared with the cisplatin group and control group （ all P 〈 0.05）. Meanwhile, no significant differences were found between salinomycin group and combination group as well as between the cisplatin group and control group （ all P〉0.05）. No significant difference was found in the expression of P70S6K and S6 between these four groups （ all P 〉 0.05 ）. Conclusion Salinomycin can reverse the cisplatin resistance of human naso- pharyngeal carcinoma cells CNI/12/DDP which is possibly mediated by improving caspases activity and inhibiting both Wnt/β-catenin signaling pathway and mTORC1 signaling pathway.

关 键 词：鼻咽癌 盐霉素 顺铂 耐药 细胞凋亡 

分 类 号：R739.6[医药卫生—肿瘤]