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目的:探讨卡介苗(BCG)对膀胱癌细胞株T739经Toll样受体介导的NF-κB p65活性影响。方法:采用SYBRGreen嵌合荧光定量Real Time-PCR,观察BCG(0.25g/L)刺激T739后不同时间点各个mRNA相对表达量(Ratio),设LPS和空白组作对照;采用转录因子结合DNA的ELISA方法检测NF-κB p65活性。结果:BCG刺激后T739细胞TLR2、CD14和MD-2mRNA表达均明显增加,Max.Ratio分别为5.3(2h),2.6(4h),7.4(2h),TLR4mRNA增加不显著,Max.Ratio为1.6(6h);BCG作用T739细胞后NF-κB p65活性显著增加,阻断TLR4后,NF-κB p65活性受到显著抑制(P<0.05);阻断TLR2后NF-κB p65活性却显著性增强(P<0.01)。结论:BCG作用T739后TLR2、CD14、MD-2和TLR4mRNA表达均不同程度增加,BCG可经TLR4使T739细胞NF-κB p65活性增强,不是经TLR2使T739细胞NF-κB p65活化。
Objective: To investigate the effect of BCG on Toll-like receptor-mediated NF-κB p65 activity in bladder cancer cell line T739. Methods: SYBRGreen fluorescence quantitative real-time PCR was used to observe the relative expression of mRNA at different time points after T739 stimulated by BCG (0.25g / L). LPS and blank control were used as control; transcription factor combined with DNA ELISA method was used to detect NF-κB p65 activity. Results: The expressions of TLR2, CD14 and MD-2 mRNA in T739 cells were significantly increased after BCG stimulation. The Max.Ratio values were 5.3 (2h), 2.6 (4h) and 7.4 (2h) (P <0.05). After blocking TLR2, the activity of NF-κB p65 was significantly increased (P <0.05) <0.01). CONCLUSION: The expressions of TLR2, CD14, MD-2 and TLR4 mRNA in BCG treated with BCG are all increased to different extents. BCG can enhance the activity of NF-κB p65 in T739 cells by TLR4, but not by TLR2.