论文部分内容阅读
目的观察NF-κB核转录因子在砷诱导人正常膀胱上皮细胞(SV-HUC-1)环氧化酶-2(COX-2)表达中的作用。方法在细胞培养液中加入不同浓度的NaAsO2,24 h后提取细胞总RNA和核蛋白,用RT-PCR和Westernblot方法分别分析COX-2 mRNA表达和NF-κB蛋白水平;选择COX-2高表达的NaAsO2处理组,再用NF-κB抑制剂(PDTC)处理,观察COX-2 mRNA表达水平的变化。结果 4、8μmol/L NaAsO2处理细胞后,COX-2 mRNA表达水平显著高于对照组(P<0.05);4μmol/L NaAsO2处理组细胞NF-κB蛋白水平也显著提高,高于对照组,而10μmol/LNaAsO2处理组细胞NF-κB蛋白水平低于对照组(P<0.05);同时用4μmol/L NaAsO2和PDTC共同处理细胞后,COX-2 mRNA表达水平降低,显著低于单纯用4μoml/L NaAsO2处理细胞的COX-2 mRNA表达水平(P<0.05)。结论低剂量的砷能够诱导人正常膀胱上皮细胞COX-2 mRNA和NF-κB蛋白表达水平的增高,砷诱导的核转录因子NF-κB活化在砷诱导的COX-2表达水平增高中起一定作用。
Objective To investigate the role of NF-κB nuclear transcription factor in arsenic-induced expression of cyclooxygenase-2 (COX-2) in human normal bladder epithelial cells (SV-HUC-1) Methods The total RNA and nuclear protein of cells were extracted with different concentration of NaAsO2 in cell culture medium for 24 h. The expression of COX-2 mRNA and NF-κB protein were detected by RT-PCR and Western blot respectively. NaAsO2 treatment group, and then treated with NF-κB inhibitor (PDTC) to observe the change of COX-2 mRNA expression level. Results The expression of COX-2 mRNA in cells treated with 4 and 8μmol / L NaAsO2 was significantly higher than that in the control group (P <0.05). The protein level of NF-κB in 4μmol / L NaAsO2 group was significantly higher than that in the control group The protein level of NF-κB in 10μmol / LNaAsO2 treated group was lower than that in control group (P <0.05). Concurrently with 4μmol / L NaAsO2 and PDTC, COX-2 mRNA expression level was significantly lower than that of 4μmol / COX-2 mRNA expression in NaAsO2-treated cells (P <0.05). Conclusion Arsenic at a low dose can induce the expression of COX-2 mRNA and NF-κB protein in human normal bladder epithelial cells. Arsenic-inducible NF-κB activation plays a role in arsenic-induced COX-2 expression .