负向调控鼻咽癌微小RNA-9表达促进紫外线介导的活性氧损伤  

Inhibition of micro RNA-9 expression promotes UV-indueed ROS damage in nasopharyngeal carcinoma cells

作　　者：郑朝攀 韩灵[2] 侯伟坚[2] 文译辉[2] 傅然[2] 马仁强[2] 文卫平[2] 

机构地区：[1]深圳市人民医院耳鼻咽喉科,518020 [2]中山大学附属第一医院耳鼻咽喉科医院

出　　处：《中华耳鼻咽喉头颈外科杂志》2013年第8期668-672,共5页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

摘　　要：目的探讨负向调控鼻咽癌细胞微小RNA-9（microRNA-9，miR-9）表达对紫外线介导的活性氧损伤的调节作用。方法应用脂质体Lipofectamine2000转染抑制鼻咽癌细胞miR-9表达，转染抑制对照剂作为参照。应用紫外线照射45min后，二氯二氢荧光素双醋酸盐检测两组细胞紫外线诱导的活性氧变化；AnnexinV—FITC法检测细胞凋亡和碱性彗星实验检测细胞DNA损伤差异；并观察谷胱甘肽在调节紫外线活性氧损伤中的作用。结果抑制CNE-2细胞miR-9表达后，miR-9抑制组和抑制对照组细胞在紫外线照射的活性氧水平平均（x±s，下同）分别为26895±218和15765±927，两者差异有统计学意义（t=39．754，P〈0．001）；紫外线照射后miR-9抑制组DNA损伤和凋亡率分别为28．0％±10．0％和8．0％±0．9％，显著大于抑制对照组的23．6％±9．2％和4．5％±0．8％：并且miR-9抑制组细胞中谷胱甘肽的表达低于抑制剂对照组，含量分别为（1．87±0．15）μmol／L和（9．85±0．15）μmol／L（t=-48．832，P〈0．001）。负向调控CNE-1细胞表达抑制了谷胱甘肽的表达和增加了紫外线介导的活性氧水平。结论鼻咽癌细胞中，负向调控miRO能够促进肿瘤细胞对紫外线介导的活性氧损伤。Objective To investigate the effects of down-regulated miR-9 expression on ultraviolet rays （UV）-induced reactive oxygen species （ROS） damage in nasopharyngeal carcinoma （NPC） cells. Methods The NPC cells were transfected with inhibitors of miR-9 by lipofectamine to decrease the expression of miR-9, and the cells transfected with inhibitor control as the control. ROS levels following UV exposure were examined with DCF-DA method and the concentration of glutathione was analyzed via the benzoic acid method; DNA damage and apoptosis also were evaluated. Results There was significant difference in ROS levels between miR-9 expression-inhibited cells and control cells （26 895 ± 218 vs 15 765± 927, t = 39. 754, P 〈 0. 001 ） , and also there were significant differences in DNA damage rates （28.0% ± 10. 0% vs 23.6% ±9. 2% ） and in apoptosis rates （8.0% ±0. 9% vs4. 5% ±0. 8% ） following UV exposure between two groups of cells. The miR-9 expression-inhibited cells showed lower level （ 1.87 ± 0. 15） μmol/L of glutathione compared with the control cells （9.85 ± 0. 15） μmol/L （t = -48. 832, P〈0.001）. Conclusion Inhibition of miR-9 expression promoted UV-induced ROS damage in nasopharyngeal carcinoma cells.

关 键 词：鼻咽肿瘤 微RNAS 活性氧 紫外线 

分 类 号：R739.63[医药卫生—肿瘤]