淫羊藿甙逆转双酚A促甲状腺癌B-CPAP细胞活性的研究  

The bisphenol A-enhanced activity of thyroid carcinoma cell line B-CPAP is inhibited by Icarrin

作　　者：郑传铭[1] 刘小珍[2] 李清林[3] 王佳峰[1] 谭卓[1] 葛明华[1] 

机构地区：[1] 浙江省肿瘤医院头颈外科,杭州310022 [2] 浙江省肿瘤医院生物标本库,杭州310022 [3] 浙江省肿瘤医院药剂科,杭州310022

出　　处：《中华耳鼻咽喉头颈外科杂志》2017年第6期458-462,共5页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

基　　金：国家自然科学基金（81550003）;浙江省自然科学基金（LY17H280003）;浙江省中医药科学研究基金（2014ZB025）

摘　　要：目的 分析淫羊藿甙（icariin,ICA）逆转环境内分泌干扰物双酚A（bisphenol A,BPA）促甲状腺癌B-CPAP细胞增殖功能及可能机制.方法 采用CCK-8试剂盒检测不同浓度ICA、BPA处理组B-CPAP细胞增殖变化,流式细胞仪技术检测细胞凋亡率及活性氧（reactive oxygen species,ROS）的表达,超氧化物歧化酶（superoxide dismutase,SOD）试剂盒检测B-CPAP胞内超氧化物歧化酶活性,脂质氧化试剂盒检测胞内丙二醛（malondialdehyd,MDA）表达变化,采用蛋白质印迹法（Westernblot）检测Bcl-2及γ-HA2X蛋白表达.采用SPSS 18.0统计学软件进行统计学分析.结果 3＆#215;10-1moL/L BPA处理48 h后,BPA组A值明显高于对照组（1.089±0.053比0.935±0.010,P＜0.05）;BPA ＋ICA25组、BPA＋ ICA50组、BPA＋ ICA1oo组、BPA＋ ICA200组A值分别为0.780 ±0.036、1.007±0.050、0.958±0.033、0.625±0.064,均高于BPA组（P值均＜0.01）.72 h各处理组增殖趋势同48 h相似,24h各组差异无统计学意义.48 h后BPA凋亡率为（19.272 ±0.186）％,而对照组为（22.412±0.238）％（P＜0.05）;BPA＋ ICAso组、BPA＋ICA200组B-CPAP细胞凋亡率分别为（23.688 ±0.412）％、（30.270±0.696）％ （P＜ 0.01）.BPA ＋ICA50组、BPA＋ ICA2m组ROS均高于BPA组（1 772±37、2 041±16比806±21,P值均＜0.01）,呈剂量依赖性.对照组和BPA组的Bcl-2蛋白表达量高于BPA ＋ICA5o和BPA＋ICA200组（7 120±151、9 801 ±286比5 902±171、4 203 ±216,P ＜0.01）.结论 BPA可良好地促进甲状腺癌B-CPAP细胞增殖、降低细胞凋亡率,而这种作用可被ICA逆转,其作用可能途径为诱导胞内ROS高表达且抑制抗氧化酶体系表达,导致细胞氧化损伤,从而诱导凋亡.Objective To investigate the effect of icariin （ICA） on the bisphenol A （BPA）enhanced proliferation function of thyroid carcinoma cell B-CPAP and underlying mechanism.Methods The proliferation of Gastric B-CPAP cell line was evaluated by cell counting kit-8 （CCK-8）.Apoptosis and ROS expression in B-CPAP cells were detected by flow cytometry.The expression of superoxide dismutase （SOD） and malondialdehyde （MDA） in B-CPAP cells were measured by individual assay kits.The expressions of Bcl-2 and γ-HA2X were detected by Western blot.SPSS 18.0 software was used to analyze the data.Results B-CPAP cell activity was promoted by treatment with 3 ＆#215; 10-7 mol/L BPA for 48 h,with significant difference in absorbance between BPA and control groups （1.089 ±0.053 vs 0.935 ±0.010,P 〈0.05）.The cell activities of BPA ＋ ICA25,BPA ＋ ICA50,BPA ＋ ICA100 and BPA ＋ ICA200 groups was 0.780 ±0.036,1.007 ±0.050,0.958 ±0.033 and 0.625 ± 0.064,respectively （all P 〈 0.01）.The proliferation of B-CPAP cells treated with BPA for 72 hours showed a similar trend to 48 hours.There was no significant difference between all treatment groups in 24 hours.The apoptosis rate was （19.272 ± 0.186）% in BPA-treated cells,and was （22.412 ± 0.238） % in control cells （P 〈 0.05）.The apoptosis rates of BPA ＋ ICA50 and BPA ＋ ICA200 groups were （23.688 ± 0.412） % and （30.270 ± 0.696） %,respectively （P 〈 0.01）.The intracellular accumulation of ROS in BPA,BPA ＋ ICA50,and BPA ＋ ICA200 groups were 806 ± 21,1 772 ± 37,2 041 ± 16,respectively （P 〈 0.01）.The expressions of anti-apoptotic protein Bcl-2 in control,BPA,BPA ＋ ICA50,BPA ＋ ICA200 groups were 7 120 ± 151,9 801 ± 286,5 902 ± 171 and 4 203 ±216,respectively （P 〈0.01）.Conclusion BPA can promote the proliferation of thyroid carcinoma B-CPAP cells and decrease the apoptosis of cells,and this effect can be inhibited by ICA.The possible mechanism is to induce high expression of intracellular ROS and inhib

关 键 词：甲状腺肿瘤 超氧化物歧化酶 淫羊藿甙 

分 类 号：R285.5[医药卫生—中药学]