RT-PCR检测人膀胱上皮细胞双功能氧化酶表达

Semi-quantitative analysis of dual oxidase expression in uroepithelial cells by RT-PCR

目的:检测人膀胱上皮细胞(T24细胞)是否表达双功能氧化酶(Duox-1、Duox-2),并观察炎症细胞因子及佛波酯(PMA)对基因表达水平的影响。方法:体外培养人膀胱上皮(T24)细胞,经佛波酯(PMA)、炎症细胞因子(TNF-α及IFN-γ)刺激后提取细胞总RNA,采用半定量RT-PCR方法观察细胞Duox-1,Duox-2 mRNA表达水平。结果:Duox-1,Duox-2 mRNA在T24细胞中组成性表达,在佛波酯(PMA)、TNF-α及IFN-γ作用下,Duox-2基因表达水平明显增加,而Duox-1基因表达水平无明显变化。结论:双功能氧化酶Duox-1和Duox-2基因在膀胱上皮细胞表达,并受炎症刺激的影响,其在膀胱上皮细胞中的生理功能值得深入研究。

Objective： To investigate the expression and regulation of dual oxidase genes （Duox-1 and Duox-2） in uroepithelial cells. Methods： Human uroepithelial cell line T24 was cultured with 1640 medium and the cells were stimulated with cytokines （TNF-α plus IFN-γ） or Pborbol-12-myristate-13-acetate （PMA）. Total RNA was isolated using the Trizol reagent and the expression level of Duox-1 and Duox-2 genes were analyzed by semi-quantitative RT-PCtL Results： There are constitutive expression of both Duox-1 and Duox-2 genes in uroepithelial cell line T24, but only the expression of Duox-2 gene was enhanced by TNF-α plus IFN-γ（26. 8 fold increase）or PMA （4. 5 fold increase）. Conclusions： The expression and regulation by inflammation media of dual oxidase genes in uroepithelial cells suggested that they may play a role in the mucosal inflammatory and innate immune response in the urinary tract.