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Screening and expression of specific SiRNA targeting IKKβ in human hypertrophic scar fibroblasts and construction of IKKβ-siRNA expression vector
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 8
- DOI:
- 10.3321/j.issn:1000-2790.2009.08.004
- Key Word:
- 瘢痕,肥大性;成纤维细胞;IKKβmRNA;IKKβsiRNA表达栽体
Abstract： 目的:体外合成并筛选人增生性瘢痕成纤维细胞特异性IKKβ-siRNA.构建其表达载体并检测其表达.方法:化学合成3条IKKβ-siRNA,分别转染体外培养的人增生性瘢痕成纤维细胞,采用RT-O-PCR筛选出特异性IKKβ-siRNA,并采用PU6质粒构建载体,RT-Q-PCR检测其表达.结果:合成的3条IKKβ-siRNA链中,仅第二条有明显抑制IKKβ的mRNA表达的作用,并通过酶切鉴定和测序结果证明成功构建IKKβ-siRNA载体,经转染靶向IKKβ-siRNA的成纤维细胞,IKKβ的表达在转染后24 h及48 h均受到抑制.结论:应用RT-PCR可筛选出针对人增生性瘢痕成纤维细胞的特异性IKKβ-siRNA;可构建具有干扰效果的IKKβ-siRNA表达载体,转染后可以降低IKKβ的表达.
- [1]马建民,赵家良,卞爱琳,等.大鼠结膜囊成纤维细胞TGFβ2特异性siRNA的筛选及其载体构建的研究[J].2006,(6).
- [2]汤华主编.RNA干扰原理与应用[M].1900.
- [3]Melisi D,Chiao PJ.NF-kappaB as a target for cancer therapy.[J].2007,11(2).
- [4]Nishikura K.Editor meets silencer: crosstalk between RNA editing and RNA interference[J].2006,7(12).
- [5]Dykxhoorn DM,Novina CD,Sharp PA.Killing the messenger: short RNAs that silence gene expression.[J].2003,4(6).
- [6]Michael J May,Sankar Ghosh.Signal transduction through NF-κB[J].1998,19(2).
- [7]Ninomiya Tsuji J,Takaesu G,Matsumoto K,etc.TAK1 is critical for IkappaB kinase-mediated activation of the NF-kappaB pathway.[J].2003,326(1).