Suppressed expression of thioredoxin-2 by small interference RNA in A549 cells exposed to hyperoxia reduced expression of nicotinamide-adenine dinucleotide dehydrogenase subunit 1 and cytochrome C oxidase Ⅰ
10.3760/cma.j.issn.1007-9408.2010.02.011
- VernacularTitle:小分子干扰RNA抑制高氧暴露下人肺腺癌A549细胞中的硫氧还蛋白-2对还原型烟酰胺腺嘌呤二核苷酸脱氢酶亚单位1、细胞色素C氧化酶工表达的影响
- Author:
Cheng CAI
;
Liwen CHANG
;
Wenbin LI
;
Yan CHEN
;
Ruiyan SHAN
;
Wei LIU
;
Rui PAN
- Publication Type:Journal Article
- Keywords:
RNA,small interfering Thioredoxins;
NADH dehydrogenase;
Electron transportcomplex IV,Hyperoxia;
Lung diseases
- From:
Chinese Journal of Perinatal Medicine
2010;13(2):128-133
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effects of expression of thioredoxin-2(Trx-2) suppressed by small interference RNA(SiRNA) in A549 cells exposed to hyperoxia on expression of nicotinamide adenine dinucleotide(NADH) dehydrogenase subunit 1(ND1)and cytochrome C oxidase Ⅰ(COX Ⅰ). Methods A549 cells were gained by serial subcultivation in vitro and transfered with synthetic Trx-2 sequence-specific SiRNA and then were randomly divided into air group without interference,hyperoxia group without interference,air group after interference,and hyperoxia group after interference.After exposure to oxygen or room air for 12,24 and 48 h,expressions of Trx-2,ND1 and COX Ⅰ mRNA of these cells were detected by reverse transcription-polymerase chain reaction (RT-PCR),and Trx-2 protein was detected by Western blot. Results (1)Sequence-specific SiRNAtargeting Trx-2 could significantly down-regulate its expression in A549 cells.(2)Trx-2 mRNA levds inhyperoxia group without interference at 24 h was higher than those in air group without interference(0.7799±0.1249 VS 0.4424±Ⅰ.1140,P<0.05).Th-2 mRNA levels in hyperoxia group after ireedcrence at 24 hand 48 h were 0.2774±0.0174 and 0.2587±0.0069,lower than those in air group after interference andhyperoxia group without interference (P<0.05).(3)ND1 mRNA levels in hyperoxia group without interference at 24 h was 0.6609±0.0368,lower than those in air group without interference(0.8898±0.1049)(P<0.05).ND1 mRNA levels in hyperoxia group after interference at 12 h was 0.8848±0.0135,higher than those in air group after imederence(P<0.05).ND1 mRNA levels in hypemxia group afterinterference at 48 h was 0.3808±0.0937,lower than those in air group after imerference and hyperoxiagroup without interference(P<0.05).(4)COXI mRNA levels in hypemxia group without inteference at 12,24 and 48 h were 1.7313±0.4331,2.1929±0.6722 and 2.0754±0.2584,higher than those in air group witheUt interference,respectively (P<0.05). Conclusions ND1 and COXⅠ participate in thedevelopment of hyperoxia indUCed lung.injury,and Trx-2 is likely to have protective effect on mitochondria ofA549 cells in hyperoxia lung injury.
