CAJ | 학술논문

背景：miRNA通过调节特异靶基因的表达，在疾病的发展及预后中起着重要作用。目的：探索miRNA-21在宫颈癌Hela细胞中对TLR4基因的调控关系。方法：通过 miRNA 靶基因预测网站寻找可能与 miRNA-21相互作用的靶基因，利用已构建的携带pri-miRNA-21基因重组腺病毒载体，包装并大量扩增病毒感染 Hela 细胞，检测荧光蛋白的表达水平，提取感染48 h蛋白，通过Western印迹检测TLR4蛋白表达。从蛋白水平验证在Hela细胞中miRNA-21与靶基因TLR4的靶向调控关系。结果与结论：100 MOI的重组腺病毒pAd/pri-miRNA-21、pAd/neg可以成功感染Hela细胞。生物信息学方法显示miRNA-21和TLR4存在可能的结合位点。发现感染pAd/pri-miRNA-21组与对照组pAd/neg及空白组相比，TLR4蛋白的表达量明显降低。证实了miRNA-21可以负调节靶基因TLR4的表达。
배경：miRNA통과조절특이파기인적표체，재질병적발전급예후중기착중요작용。목적：탐색miRNA-21재궁경암Hela세포중대TLR4기인적조공관계。방법：통과 miRNA 파기인예측망참심조가능여 miRNA-21상호작용적파기인，이용이구건적휴대pri-miRNA-21기인중조선병독재체，포장병대량확증병독감염 Hela 세포，검측형광단백적표체수평，제취감염48 h단백，통과Western인적검측TLR4단백표체。종단백수평험증재Hela세포중miRNA-21여파기인TLR4적파향조공관계。결과여결론：100 MOI적중조선병독pAd/pri-miRNA-21、pAd/neg가이성공감염Hela세포。생물신식학방법현시miRNA-21화TLR4존재가능적결합위점。발현감염pAd/pri-miRNA-21조여대조조pAd/neg급공백조상비，TLR4단백적표체량명현강저。증실료miRNA-21가이부조절파기인TLR4적표체。
BACKGROUND:MicroRNAs (miRNA) through regulating specific target gene mRNA expression play important roles in different processes of diseases. OBJECTIVE:To study the interaction of miRNA-21 with its target gene TLR4 in Hela cels. METHODS:The candidate target gene of miRNA-21 was determined according to miRNA analysis databases. The constructed recombinant adenovirus vector carrying pri-miRNA-21 gene was used, which could package and amplify viruses to transfect Hela cels. Then, the expression of fluorescent proteins was detected. Forty-eight hours after transfection of miRNA-21 or control, extracted proteins were used for detection of TLR4 protein using western blot assay. RESULTS AND CONCLUSION:Recombinant adenoviruses pAd/pri-miRNA-21 and pAd/neg at 100 MOI could successfuly infect Hela cels. Bioinformatic analysis suggested several possible binding sites between miRNA-21 and TLR4. The experimental results showed that miRNA-21 down-regulated TLR4 at protein levels, indicating that miRNA-21 can interfere with the expression of TLR4 target gene.