摘要
目的观察Slug在TGF-β1诱导的人肾小管上皮细胞向间质细胞转分化中的表达及对黏附分子E-cadherin表达的影响。方法构建能表达针对Slug的小干扰RNA(Small interferingRNA,siRNA)的干扰载体(SlugsiRNAvector)和表达不针对任何已知mRNA的siRNA的阴性对照RNA干扰载体(Control siRNA vector),并用脂质体转染法瞬时转染HKC细胞。用TGF-β1分别处理HKC、siSlug和siC,刺激48h后,分别采用PT-PCR和Western blot技术检测各组Slug、α-SMA(alpha-smooth muscleactin)、和E-cadherin的表达水平。结果 TGF-β1刺激后Slug表达水平增高,E-cadherin表达水平降低;通过siRNA干扰后Slug表达减弱,而E-cadherin的表达增强。结论在TGF-β1诱导的HKC细胞EMT过程中,Slug表达与E-cadherin呈负相关,且RNAi阻滞Slug表达能有效地抑制TGF-β1诱导EMT,提示Slug是介导TGF-β1有效地诱导EMT的重要分子。
To investigate the expression of slug in human renal tubular epithelial-mesenchymal transdifferentiation（EMT） induced by transforming growth factor-β1（TGF-β1） in vitro,and the inhibitory effects of slug on E-cadherin,RNA interference plasmid that can express small interfering RNA（siRNA） targeting Slug（Slug siRNA vector） or express siRNA that does not match any known human coding mRNA（control siRNA vector） was designed,constructed,and transiently lipotransfected transient.into HKC cell line.HKC,HKC-siSlug and HKC-siC were treated with TGF-β1 for 48 hours,and then the levels of Slug,α-SMA（alpha-smooth muscle actin）,and E-cadherin expression were determined by reverse transcription-poly-merase chain reactin（RT-PCR） and Western blotting.The results showed that the expression of Slug was increased but the expression of E-cadherin was decreased after the treatment of TGF-β1;Slug expression was decreased obviously but E-cadherin expression was increased through the siRNA interference.The results indicate that in HKC cell EMT induced by TGF-β1,there is a negative correlation between the Slug expression and E-cadherin expression,and RNAi targeting Slug can efficiently inhibit EMT,which imply that Slug is essential to EMT induced by TGF-β1.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2010年第10期872-875,882,共5页
Immunological Journal
