摘要
目的探讨细胞因子白介素-1β(IL-1β)对人肺腺癌细胞株A549细胞环氧合酶2(COX-2)表达的影响及调节机制。方法A549细胞培养达80%融合后,用不同浓度的IL-1β(0、0.1、1、5和10ng/mL)干预或用5ng/mLIL-1β干预不同时间(0、3、6、9、12和24h)观察白介素-1β(IL-1β)对A549细胞COX-2表达的影响;用5ng/mLIL-1β与有丝分裂原激活的细胞外调节激酶(ERK)抑制剂PD098059、P38有丝分裂原激活的蛋白激酶(MAPK)抑制剂SB203580、蛋白激酶C(PKC)抑制剂H-7共同干预A549细胞观察IL-1β诱导A549细胞COX-2表达的信号传导途径;用COX-2系列报告基因质粒及对照质粒转染A549细胞后予以IL-1β干预,以不加IL-1β干预的细胞为对照组观察IL-1β对COX-2启动子的影响;Westernblot检测COX-2蛋白表达,RT-PCR检测COX-2mRNA表达。COX-2报告基因活性应用荧光素酶活性分析法测定。结果A549细胞COX-2蛋白表达和mRNA表达呈IL-1β浓度依赖性表达增加,干预9h表达达高峰;SB203580、H-7可明显抑制IL-1β诱导A549细胞COX-2表达,PD098059对IL-1β诱导A549细胞COX-2表达无影响;IL-1β对COX-2报告基因无影响。结论PKC途径和P38MAPK途径参与IL-1β诱导A549细胞COX-2蛋白表达。
[Objective] To investigate the effect of IL-1β on COX-2 expression and its signal pathways in A549 cells. [Methods] A549 cells were chosen to be stimulated with IL-1β to test the effect of IL-1β on COX-2 expression, then to be stimulated with H-7, a protein kinase C inhibitor, SB203580, a P38 mitogen activated protein（MAP） kinase inhibitor and PD098059, a mitogen activated and extraeellular regulated kinase （MEK1） inhibitor with or without IL-1β respectively. COX-2 mRNA and protein were determined by RT-PCR and Western blot respectively. [Results] RT-PCR and Western blot showed that IL-1β induced COX-2 mRNA and protein expression in a timeand dose-dependent manner in A549 cells. SB203580 and H-7 abrogated COX-2 expression induced by IL-1β, while PD098059 had no effect on it. [Conclusions] Signal pathways PKC and P38 MAPK may be involved in the IL-1β- induced COX-2 expression in A549 cells.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2006年第16期2405-2409,共5页
China Journal of Modern Medicine
基金
It is supported by a grant from Hunan Science and Technology Department (01SSY2008- 38).
