肺瘤平膏对共培养条件下A549细胞NF-κB通路mRNA及蛋白表达的影响  

Effects of Feiliuping Plaster on NF-κB related gene and protein expression in A549 cell under co-culture conditions

作　　者：陈赐慧[1] 李卫东[2] 刘瑞[2] 花宝金[2] 

机构地区：[1]浙江省中医院,杭州310006 [2]中国中医科学院广安门医院,北京100053

出　　处：《中华中医药杂志》2016年第4期1207-1210,共4页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家科技部重大新药创制专项课题(No.2010ZX09102-216);国家自然科学基金面上项目(No.81273718);国家自然科学基金青年基金项目(No.81102587)~~

摘　　要：目的:观察肺瘤平膏对共培养条件下人肺腺瘤细胞(A549细胞)核因子-κB(NF-κB)通路mRNA及蛋白表达的影响。方法:制备肺瘤平膏含药血清;建立A549细胞与巨噬细胞的共培养体系;通过荧光定量PCR和Western blot观察肺瘤平膏对共培养条件下A549细胞NF-κB通路mRNA及蛋白表达的影响。结果:共培养条件下,A549细胞变成间充质细胞样,部分可被肺瘤平膏所逆转。共培养条件下,A549细胞中CHUK、IKBKB、NFKBIA、RelA mRNA表达量明显升高(P<0.05);干预后,除NFKBIA外,CHUK、IKBKB、RelA表达量均显著降低(P<0.05)。而A549细胞总蛋白中NF-κB p65、p-IKKα/β、p-IκBα的表达量升高(P<0.05),IKKβ、IκBα的表达量降低(P<0.05),IKKα表达量无明显变化;干预后可降低IKKα、p-IKKα/β、p-IκBα的表达量(P<0.05),升高IκBα的表达量(P<0.05)。胞浆和胞核蛋白中p-NF-κB p65的含量亦明显升高(P<0.05),干预后可降低此其表达量(P<0.05),并且降低其胞核/胞浆比例。结论:肺瘤平膏可以通过抑制CHUK基因的表达,并且减少IKKα/β的磷酸化,抑制其激活,从而抑制IκBα的磷酸化,避免IκBα的磷酸化带来的NF-κB p65核转位。Objective： To observe the effects of Feiliuping Plaster on NF-κB related gene and protein expression in A549 cell under co-culture conditions. Methods： Feiliuping Plaster-containing serum（FLP） is prepared, and co-culture system of A549 cells and macrophage is established. The influence of Feiliuping Plaster on NF-κB related gene and protein expression in A549 cell under co-culture conditions is observed by using RT-PCR and Western blot techniques. Results： A549 cells develop into mesenchymal-like cells in co-culture conditions, which can be reversed by FLP partly. Under co-culture conditions, the mRNA expression of CHUK, IKBKB, NFKBIA, and RelA significantly increased（P〈0.05）, while the expression of all the above indexes expect NFKBIA decreased significantly after intervening（P〈0.05）. The total protein expression of NF-κB p65, p-IKKα/β and p-IκBα increased in A549 cells（P〈0.05）, the expression of IKKα and IκBα protein decreased（P〈0.05）, and the expression of IKKα has no significant change. After intervening, the expression of IKKα, p-IKKα/β, and p-IκBα reduced（P〈0.05）, and the expression of IκBα increased significantly（P〈0.05）. The contents of p-NF-κB p65 in cytoplasm and nucleus also increased obviously（P〈0.05）, but the expression（P〈0.05） and the nucleus/cytoplasm ratio of the two proteins can be reduced by intervening with FLP. Conclusion： FLP can inhibit the phosphorylation of IκBα to avoid NF-κB p65 nuclear translocation caused by IκBαphosphorylation by inhibitting the expression of CHUK, reducing the phosphorylation of IKKα/β, and preventing its activation.

关 键 词：肺瘤平膏 肺癌 A549细胞 共培养 核因子-ΚB 

分 类 号：R273[医药卫生—中西医结合]