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
出处
《中华耳鼻咽喉头颈外科杂志》
CAS
CSCD
北大核心
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）
关键词
甲状腺肿瘤
超氧化物歧化酶
淫羊藿甙
Thyroid neoplasms
Superoxide dismutase
Bisphenol A
Icarrin
