PEITC对人鼻咽癌CNE-1/CNE-2细胞凋亡作用及机制研究  

Role and Mechanism of Phenethyl Isothiocyanate on Apoptosis of Nasopharyngeal Carcinoma Cell Line CNE-1/2

作　　者：张功学[1] 王明华[1] 李道坤[2] 

机构地区：[1]湖北文理学院附属枣阳医院病理科,湖北枣阳441200 [2]湖北文理学院分子医学中心,湖北襄阳441053

出　　处：《湖北文理学院学报》2016年第5期78-84,共7页Journal of Hubei University of Arts and Science

基　　金：襄阳市科技局项目(襄科业[2013]66号)

摘　　要：目的:观察PEITC(异硫氰酸苯乙酯)对人鼻咽癌CNE-1/2细胞生长的影响以及对正常鼻咽部上皮细胞的毒性作用,探讨其抗肿瘤作用机制.方法:MTT法测定PEITC对CNE-1/2细胞生长的抑制作用;流式细胞仪检测PEITC对CNE-1/2细胞凋亡和氧化还原水平的影响;NAC阻断后其氧化还原水平的变化.结果:PEITC能抑制CNE-1/2细胞生长,随时间延长和剂量增加肿瘤细胞数明显减少,CNE-2凋亡作用更加明显,而对NP-69细胞组细胞毒性较低.当10.0 mg·ml^(-1) PEITC在96 h时间点,NP-69细胞组IC50(12.62±0.52)与CNE-1组(3.57±0.34)、CNE-2细胞组IC_(50)(2.64±0.20)差异具有统计学意义(P<0.01);用Annexin V/PI染色流式细胞检测各组细胞经PEITC处理24 h后细胞发生凋亡,随着PEITC浓度(0、5.0、10.0 mg·ml^(-1))的增加,细胞凋亡率增加;当PEITC10.0 mg·ml^(-1)浓度时,CNE-1/CNE-2凋亡率与对照组差异具有统计学意义(P<0.01);DCFH-DA探针标记后流式检测活性氧水平结果显示:CNE-1/CNE-2活性氧水平分别在6、12h时间点与对照组差异具有统计学意义(P<0.01);Rhodamine-123试剂盒测定线粒体膜电位显示:10.0 mg·ml^(-1)浓度PEITC时CNE-1/CNE-2线粒体膜电位在24 h时间点与对照组差异具有统计学意义(P<0.01);用NAC预处理各组细胞后检测细胞活性氧及线粒体膜电位均出现反向作用.结论:PEITC能抑制人鼻咽癌CNE-1/2细胞的生长,同时对NP-69细胞具有低毒性,其抗肿瘤作用可能与其干扰细胞内的氧化还原反应,诱导细胞凋亡有关.Objective： to investigate the effect and mechanism of phenethyl isothiocyanate（PEITC） on apoptosis of nasopharyngeal carcinoma CNE-1/2 cell, and the influence of the toxic effects of normal nasopharyngeal epithelial cells, to discuss its antitumor mechanism. Methods： determination of PEITC CNE-1/2 cell growth inhibition by MTT method; detection of PEITC on CNE-1/2 cell apoptosis and the influence of oxidation reduction level by flow cytometry instrument; the change of oxidation reduction level after NAC is blocked. Results： PEITC can inhibit the growth of CNE- 1/2 cell： with the time extension and the dose increase, tumor cells significantly reduced, CNE-2 apoptosis effect become more apparent, with low cytotoxicity to NP- 69 cell group. After treated 96 h with 10.0 mg. ml^（-1） PEITC, the IC_（50）（12.62±0.52） of NP- 69 cell group, the IC50（3.57±0.34） of CNE-1 group, the IC50（2.64±0.20） of CNE-2 cell groups are statistically significant different（P 〈0.01）; Annexin V/PI staining flow detection PEITC effect after 24 h after cells apoptosis, as PEITC concentration（0, 5.0, 10.0 mg. ml^（-1）） increased, cell apoptosis rate increased; When PEITC 10.0 mg. ml^（-1） concentration, CNE-1/2 apoptosis rate and the control group differences statistically significant（P 〈0.01）; DCFH- DA probe results showed that current detecting reactive oxygen levels, respectively at 6 and 12 h time points and the control group differences statistically significant（P 〈0.01）; Rhodamine-123 kit determination of mitochondrial membrane potential shows that： when the concentration of 10.0 mg. ml^（-1）, CNE- 1/2 of mitochondrial membrane potential in 24 h time point and the control group differences statistically significant（P 〈0.01）; With NAC pretreatment groups of cells after testing reactive oxygen species and mitochondrial membrane potential in reverse. Conclusion： PEITC can inhibit the growth of nasopharyngeal carcinoma CNE-1/2 cells, has low toxicity to the cells of NP-

关 键 词：异硫氰酸苯乙酯 鼻咽癌 细胞凋亡 

分 类 号：R361.3[医药卫生—病理学]