中华耳鼻咽喉头颈外科杂志
中華耳鼻嚥喉頭頸外科雜誌
중화이비인후두경외과잡지
CHINESE JOURNAL OF OTORHINOLARYNGOLOGY HEAD AND NECK SURGERY
2013年
8期
668-672
,共5页
郑朝攀%韩灵%侯伟坚%文译辉%傅然%马仁强%文卫平
鄭朝攀%韓靈%侯偉堅%文譯輝%傅然%馬仁彊%文衛平
정조반%한령%후위견%문역휘%부연%마인강%문위평
鼻咽肿瘤%微RNAs%活性氧%紫外线
鼻嚥腫瘤%微RNAs%活性氧%紫外線
비인종류%미RNAs%활성양%자외선
Nasopharyngeal neoplasms%MicroRNAs%Reactive oxygen species%Ultraviolet rays
目的 探讨负向调控鼻咽癌细胞微小RNA-9(micro RNA-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Lipofectamine 2000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45 min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；Annexin V-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26 895 ±218和15 765±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miR-9能够促进肿瘤细胞对紫外线介导的活性氧损伤.
目的 探討負嚮調控鼻嚥癌細胞微小RNA-9(micro RNA-9,miR-9)錶達對紫外線介導的活性氧損傷的調節作用.方法 應用脂質體Lipofectamine 2000轉染抑製鼻嚥癌細胞miR-9錶達,轉染抑製對照劑作為參照.應用紫外線照射45 min後,二氯二氫熒光素雙醋痠鹽檢測兩組細胞紫外線誘導的活性氧變化；Annexin V-FITC法檢測細胞凋亡和堿性彗星實驗檢測細胞DNA損傷差異；併觀察穀胱甘肽在調節紫外線活性氧損傷中的作用.結果 抑製CNE-2細胞miR-9錶達後,miR-9抑製組和抑製對照組細胞在紫外線照射的活性氧水平平均(-x±s,下同)分彆為26 895 ±218和15 765±927,兩者差異有統計學意義(t=39.754,P＜0.001)；紫外線照射後miR-9抑製組DNA損傷和凋亡率分彆為28.0％±10.0％和8.0％±0.9％,顯著大于抑製對照組的23.6％±9.2％和4.5％±0.8％;併且miR-9抑製組細胞中穀胱甘肽的錶達低于抑製劑對照組,含量分彆為(1.87±0.15) μmol/L和(9.85 ±0.15) μmol/L(t=-48.832,P＜0.001).負嚮調控CNE-1細胞錶達抑製瞭穀胱甘肽的錶達和增加瞭紫外線介導的活性氧水平.結論 鼻嚥癌細胞中,負嚮調控miR-9能夠促進腫瘤細胞對紫外線介導的活性氧損傷.
목적 탐토부향조공비인암세포미소RNA-9(micro RNA-9,miR-9)표체대자외선개도적활성양손상적조절작용.방법 응용지질체Lipofectamine 2000전염억제비인암세포miR-9표체,전염억제대조제작위삼조.응용자외선조사45 min후,이록이경형광소쌍작산염검측량조세포자외선유도적활성양변화；Annexin V-FITC법검측세포조망화감성혜성실험검측세포DNA손상차이；병관찰곡광감태재조절자외선활성양손상중적작용.결과 억제CNE-2세포miR-9표체후,miR-9억제조화억제대조조세포재자외선조사적활성양수평평균(-x±s,하동)분별위26 895 ±218화15 765±927,량자차이유통계학의의(t=39.754,P＜0.001)；자외선조사후miR-9억제조DNA손상화조망솔분별위28.0％±10.0％화8.0％±0.9％,현저대우억제대조조적23.6％±9.2％화4.5％±0.8％;병차miR-9억제조세포중곡광감태적표체저우억제제대조조,함량분별위(1.87±0.15) μmol/L화(9.85 ±0.15) μmol/L(t=-48.832,P＜0.001).부향조공CNE-1세포표체억제료곡광감태적표체화증가료자외선개도적활성양수평.결론 비인암세포중,부향조공miR-9능구촉진종류세포대자외선개도적활성양손상.
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％ vs 4.5％ ±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.
