肺岩宁方对肿瘤坏死因子α诱导的人肺癌细胞系核转录因子κB基因表达的影响(英文)  

Effects of Feiyanning Decoction on gene expression of nuclear factor-κB activated by tumor necrosis factor-α in lung adenocarcinoma cell line

作　　者：王菊勇[1] 金叶[2] 徐振晔[1] 郑展[1] 

机构地区：[1]上海中医药大学龙华医院中医肿瘤研究所,上海200032 [2]上海大学生命科学学院生物工程系,上海200444

出　　处：《中西医结合学报》2009年第3期249-254,共6页Journal of Chinese Integrative Medicine

基　　金：上海市浦江人才计划项目(No.07pj14079);上海市重点优势学科建设项目(No.Y0302)

摘　　要：目的:探讨中药肺岩宁方含药血清对肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)诱导的人肺癌细胞系A549核转录因子κB(nuclear factor-κB,NF-κB)基因表达的影响。方法:制备肺岩宁方含药血清,以肺癌细胞系A549为靶细胞,确定肺岩宁方含药血清用药浓度。将A549细胞分为对照血清组、TNF-α组、肺岩宁方含药血清组和综合治疗组。观察用药前后A549细胞大体形态变化;四唑单钠盐法测定肺岩宁方含药血清对细胞增殖的影响;蛋白印迹法检测肺岩宁方含药血清对TNF-α诱导的细胞核NF-κB及抑制性核转录因子(inhibitorκBα,IκBα)表达的影响;将人NF-κB重组质粒用脂质体转染法导入A549细胞24h后,用肺岩宁方含药血清治疗24h,采用双荧光素酶报告基因系统测定肺岩宁方含药血清对TNF-α诱导的细胞核NF-κBp65转录活性的影响。结果:与对照血清比较,15%和20%肺岩宁方含药血清对A549细胞具有明显的增殖抑制作用(P<0.05,P<0.01)。1μg/LTNF-α能够明显提高NF-κB基因的表达,与肺岩宁方含药血清共同作用后,能够显著抑制它的表达(P<0.01),而细胞内IκBα的表达差异无统计学意义(P<0.05);肺岩宁方含药血清能够显著降低A549细胞经NF-κB重组质粒转染后的转录活性(P<0.01)。结论:肺岩宁方含药血清可抑制人肺癌细胞系增殖,可能是通过对TNF-α诱导的NF-κB基因活性的抑制作用实现的。Objective： To study the effects of Feiyanning Decoction, a compound traditional Chinese herbal medicine, on gene expression of nuclear factor-κB （NF-κB） activated by tumor necrosis factor-α （TNF-α） in lung adenocarcinoma cell line （A549）. Methods= A549 cells were incubated with rat serum containing Feiyanning at different concentrations for 24 and 48 h, respectively. Morphology of cells was observed by an inverted microscope after treatment with reagents. The cell proliferation was examined by 2-（2-methoxy-4-nitrophenyl）-3-（4-nitrophenyl）-5-（2, 4- disulfophenyl）-2H-tetrazolium, monosodium salt （WST-8） assay. The expressions of NF-κB and inhibitor κBα （1κBα） were studied by Western blotting. NF-κB-dependent luciferase reporter （3 × κB-luc） was transfected for 24 h, and the cells were treated with the reagents for 24 h, and then the transcriptional activity of NF-κB promoter was detected by luciferase assay. Results： TNF-α （1 μg/L） strongly induced the expression of NF-κB by approximately 1.76-fold compared with the control in the nuclei of A549 cells, and the induced NF-κB expression was significantly suppressed by addition of Feiyanning （P〈0.01）. In addition, Feiyanning inhibited the transcriptional activity of the NF-κB promoter. However, we observed no significant changes in IκBα. expression （P〈0.05）. Conclusion： Feiyanning Decoction can markedly inhibit human lung cancer A549 cell proliferation, which may be partly due to inhibition of NF-κB activation induced by TNF-α. It is therefore expected to be a new strategy for treating lung cancer.

关 键 词：肺肿瘤 肺岩宁方 NF—κB 肿瘤坏死因子Α 

分 类 号：R734.2[医药卫生—肿瘤]