TNF-α对人lrg基因表达的调控

Effects of TNF-α on lrg expression in human cell lines

目的:观察TNF-α对脂多糖应答基因(lrg)在人HEK293和U937细胞中表达的影响.方法:正常培养人胚肾细胞(HEK293)和人单核细胞(U937),用TNF-α(终浓度1×106U/L)刺激2h.提取刺激前后HEK293和U937细胞的总蛋白,用纯化后的兔抗人Lrg抗血清作一抗(1∶1000),对TNF-α刺激前后的HEK293和U937细胞进行Western Blot分析.提取刺激前后HEK293和U937细胞的总RNA,用RT-PCR分析TNF-α对lrg在细胞中表达的影响.以β-actin为内参.结果:Western Blot分析显示,用TNF-α刺激2h后,lrg在人HEK293和U937细胞内的蛋白含量明显上升;RT-PCR结果显示,用TNF-α刺激2h后,人HEK293和U937细胞内的lrg mRNA水平明显上升.结论:TNF-α的刺激增强了人HEK293和U937细胞内lrg的表达,提示lrg可能参与了TNF-α诱导的炎症反应.

AIM：To examine the effect of tumor necrosis factor α （TNF-α） on lipopolysaccharide response gene （lrg） expression in human cell lines HEK293 and U937. METHODS： Human embryonic cell line HEK293 and leukemia cell line U937 were cultured in RPMI 1640 with 100 mL/L fetal serum under 50 mL/L CO2 condition. The cells were treated with TNF-α at the concentration of 1 × 10^6 U/L or placebo for 2 h. Western Blot and RT-PCR were performed to detect the lrg expression. β-actin was applied as an internal control. RESULTS： The results of Western Blot indicated that the lrg expression in both HEK293 and U937 cells at protein levels were up-regulated significantly after stimulation by TNF-α for 2 h. The results of RT-PCR indicated that the lrg mRNA levels in both HEK293 and U937 cells increased after stimulation by TNF-α for 2 h. CONCLUSION ： The lrg expression in human HEK293 and U937 cells is both up-graded remarkably by the stimulation of TNF-α, which suggests that lrg probably plays roles in the inflammation induced by TNF-α.