siRNA靶向干扰组蛋白H3表达对人鼻咽癌细胞增殖的抑制作用  

Small interfering RNA-mediated silencing of histone H3 inhibits proliferation of human nasopharyngeal carcinoma cells

作　　者：李彬彬[1] 黄国良[2] 孔霞[1] 李蓉[1] 董子明[3] 何志巍[1,2] 

机构地区：[1]广东医学院病理生理学教研室,广东东莞523808 [2]广东医学院广东省医学分子诊断重点实验室,广东东莞523808 [3]郑州大学基础医学院病理生理学教研室,河南郑州450001

出　　处：《中国病理生理杂志》2013年第9期1625-1630,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81071638);广东省医学科学技术研究基金资助项目(No.A2011424)

摘　　要：目的:应用RNA干扰技术抑制人鼻咽癌细胞株CNE1中组蛋白H3的表达,观察其对鼻咽癌细胞增殖的影响。方法:构建针对组蛋白H3的小干扰RNA(siRNA)的真核表达载体,转染CNE1细胞并筛选获得稳定表达细胞株。实时荧光定量RT-PCR和Western blotting检测细胞中组蛋白H3 mRNA和蛋白表达的改变;CCK-8法和平板克隆形成实验观察细胞增殖能力的改变;双萤光素酶报告系统检测细胞激活蛋白1(AP-1)转录活性的改变。结果:与阴性对照组和空白对照组相比,siRNA-H3质粒转染组的组蛋白H3 mRNA和蛋白表达均显著下降,细胞生长速度明显减慢,表皮生长因子诱导的细胞克隆形成能力和AP-1转录活性均受到明显抑制。结论:通过RNA干扰技术阻断组蛋白H3的表达,可抑制CNE1细胞的生长、增殖,其机制可能与下调AP-1转录活性有关。组蛋白H3可能是潜在的肿瘤治疗新靶点。AIM：To study the effect of histone H3 down-regulation by RNA interference on the proliferation of human nasopharyngeal carcinoma CNE1 cells. METHODS：The small interfering RNA （siRNA） vector targeting histone H3 was constructed and transfected into CNE1 cells, and then stably transfected CNE1 cells were established. The expression of histone H3 mRNA and protein was measured by real-time fluorescence quantitative RT-PCR and Western blotting, respectively. The proliferation ability of CNE1 cells was evaluated by CCK-8 and colony-forming assays. The transcriptional activity of activator protein-1 （AP-1） was examined by dual-luciferase reporter gene assay. RESULTS：Compared with negative control and blank control groups, histone H3 mRNA and protein expression was markedly decreased in siRNA-H3 stably transfected CNE1 cells. Knockdown of histone H3 caused a significant inhibition of cell proliferation. Furthermore, the colony formation and AP-1 transcriptional activity promoted by epidermal growth factor were obviously suppressed. CONCLUSION：Knockdown of histone H3 could significantly inhibit the proliferation of CNE1 cells by down-regulating the AP-1 transcriptional activity, and therefore histone H3 might serve as a therapeutic target in cancer treatment.

关 键 词：鼻咽肿瘤 组蛋白H3 RNA干扰 激活蛋白1 

分 类 号：R739.6[医药卫生—肿瘤]