Abstract
Objective To study the mRNA expression of thioredoxirc2 suppressed by small intederence RNA(siRNA)in A549 cells exposed to hyperoxia and the relationship with cell apoptosis.To investigate the protection effect of thioredoxin-2 Trx-2 on mitochondria under hyperoxia induced lung injury.Methods A549 cell line was cultured and transfected with synthetic Trx-2-sequence-specific siRNA by Lipofectamine 2000.These cells were randomly divided into three groups as air group with interference,hyperoxia group without interference and hyperoxia group with interference.After exposure to oxygen orroom air for 12,24 and 48 h,Trx-2 and Cytc mRNA were detected by reverse transcription polymerase chain reaction.Flow cytometry was used to detect the apoptosis of A549 cells in the three groups.Results (1)Trx-2-sequence-specific SiRNA significantly down-regulated the Trx-2 mRNA expression in A549cells.(2)Trx-2 mRNA levels in hyperoxia group with interference at 24 h and 48 h(0.2774士O.0174 andO.2587±0.0069)were lower than those in hyperoxia group without interference(0.7799±0.1249 and 0.4050±0.0842)(P＜0.05).Trx-2 tuRNA in hyperoxia group without interference at 24 h was higher than that in air group with interference(0.4046±0.0161)(P＜0.05).(3)Cytc mRNA in hyperoxia group without interference at 24 h and 48 h(0.3957±0.0903 and 0.3438±0.0092)were lower than those in hyperoxia group with interference(0.6620±0.0395 and 0.4806±0.0977)and air group with interference (0.5197±0.0377 and 0.4902±0.1079)(P＜0.05).(4)The apoptosis rate of A549 cells in hyperoxia group with interference was significantly higher than that in air group with interference(P＜0.05).Conclusions Sequence-specific SiRNA targeting Trx-2 significantly dowrcregulated its expression in A549 cells;Exposure to high concentration of oxygen Can significantly change the mRNA expression of Trx-2 and Cytc,which indicates that Trx-2 may protect mitochondfia of A549 cells from hyperoxia. Key words: RNA,small interfering; Hyperoxia; Tumor cells,cultured; Lung diseases; Thioredoxin
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