Shen Wenjun, Chen Gong, Dong Rui, Zhao Rui, Zheng Shan
Department of Pediatric Surgery, Children's Hospital of Fudan University, Shanghai 201102, China.
J Pediatr Surg. 2014 Dec;49(12):1738-41. doi: 10.1016/j.jpedsurg.2014.09.009. Epub 2014 Nov 13.
MicroRNAs (miRNAs) are short, noncoding RNA molecules that act as post-transcriptional negative regulators of target mRNAs. Increasing evidence suggests that miRNAs are involved in liver fibrotic processes. Biliary atresia (BA) is characterized by rapid and progressive liver fibrosis. Therefore, we investigated the role of miRNA-21in the pathogenesis of BA.
We collected liver samples from patients with BA or liver trauma to examine the role of miRNA-21. We examined RNA expression of miRNA-21, phosphatase and tensin homolog deleted on chromosome ten (PTEN), and α-smooth muscle actin (α-SMA) in liver tissue using real-time fluorescence quantitative PCR. Western blot analyses and immunohistochemical staining were performed to evaluate protein expression of PTEN, α-SMA, and phosphorylated AKT in liver.
We found that miRNA-21was upregulated in liver samples from BA patients, whereas PTEN negatively correlated with suppression of the 3'-untranslated region (3'-UTR). Activation of the downstream AKT pathway provoked liver fibrosis by enhancing α-SMA levels.
The miRNA-21/PTEN/AKT axis promotes the fibrosis process in BA, which might be a potential therapeutic target to improve the prognosis of patients with BA.
微小RNA(miRNA)是短链非编码RNA分子,作为靶mRNA的转录后负调控因子发挥作用。越来越多的证据表明,miRNA参与肝脏纤维化过程。胆道闭锁(BA)的特征是快速进展性肝纤维化。因此,我们研究了miRNA-21在BA发病机制中的作用。
我们收集了BA患者或肝外伤患者的肝脏样本,以研究miRNA-21的作用。我们使用实时荧光定量PCR检测肝组织中miRNA-21、第10号染色体上缺失的磷酸酶和张力蛋白同源物(PTEN)以及α-平滑肌肌动蛋白(α-SMA)的RNA表达。进行蛋白质免疫印迹分析和免疫组织化学染色,以评估肝脏中PTEN、α-SMA和磷酸化AKT的蛋白表达。
我们发现BA患者肝脏样本中miRNA-21上调,而PTEN与3'-非翻译区(3'-UTR)的抑制呈负相关。下游AKT通路的激活通过提高α-SMA水平引发肝纤维化。
miRNA-21/PTEN/AKT轴促进BA的纤维化进程,这可能是改善BA患者预后的潜在治疗靶点。