Upregulation of miRNA-143,-145,-192, and-194 in esophageal epithelial cells upon acidic bile salt stimulation
Upregulation of miRNA-143,-145,-192, and-194 in esophageal epithelial cells upon acidic bile salt stimulation
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DOI：
10.1111/dote.12112
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发表时间：
2014-08-01
影响因子：
2.6
通讯作者：
van Baal, J. W. P. M.
中科院分区：
文献类型：
--
作者：
Bus, P.;Siersema, P. D.;van Baal, J. W. P. M.
巴雷特食管（BE）是一种因慢性胃食管反流而发生的食管远端化生疾病。先前的研究已经确定了be特异性microRNAs （miRNAs）与正常鳞状上皮（SQ）的比较。我们假设be特异性mirna可以通过暴露于酸和/或胆汁盐在食管SQ细胞中诱导。我们的目的是确定在酸和/或胆盐环境下，be特异性miRNAs是否在食管SQ细胞系（Het-1A）中上调，以及这是否依赖于核因子κ B （nf - κ B）。在ht - 1a细胞中进行酸和/或胆汁盐培养。实验分别在抑制或不抑制NF-kappa B通路的情况下进行。定量逆转录酶聚合酶链反应检测miRNA-143、-145、-192、-194、环加氧酶-2 （COX2）、粘蛋白2 （MUC2）和性别决定区Y-box 9的表达。为了验证，我们在反流性食管炎和正常SQ患者的活检中测定了这些mirna的水平。在酸性胆汁盐孵育过程中，miRNA-143（2.7倍）、-145（2.6倍）、-192（2.0倍）、-194（2.2倍）、COX2、MUC2和性别决定区Y-box 9的表达水平均显著升高，而单独在酸性或胆汁盐孵育过程中则无显著升高。NF-kappa B通路抑制显著降低miRNA-143、-192、-194、COX2和MUC2的表达。此外，与正常SQ相比，反流性食管炎活检中miRNA-143、-145和-194的表达增加，但miRNA-192的表达未见变化。我们的研究结果表明，酸性胆汁对be特异性mirna的上调可能是SQ向be转变的早期事件，它们的表达部分受到nf - κ B途径的调节。
Barrett's esophagus (BE) is a metaplastic condition of the distal esophagus that occurs because of chronic gastroesophageal reflux. Previous studies have identified BE-specific microRNAs (miRNAs) in comparison with normal squamous epithelium (SQ). We hypothesized that BE-specific miRNAs could be induced in esophageal SQ cells by exposure to acid and/or bile salts. We aimed to determine whether BE-specific miRNAs are upregulated in an esophageal SQ cell line (Het-1A) in an environment with acid and/or bile salts and whether this is nuclear factor-kappa B (NF-kappa B) dependent. Acid and/or bile salt incubations were performed in Het-1A cells. Experiments were performed with or without inhibiting the NF-kappa B pathway. Quantitative reverse transcriptase polymerase chain reaction was performed to determine expression of miRNA-143, -145, -192, -194, cyclo-oxygenase-2 (COX2), mucin 2 (MUC2), and sex determining region Y-box 9. For validation, we determined levels of these miRNAs in biopsies from patients with reflux esophagitis and normal SQ. Significantly increased expression levels of miRNA-143 (2.7-fold), -145 (2.6-fold), -192 (2.0-fold), -194 (2.2-fold), COX2, MUC2, and sex determining region Y-box 9 were found upon acidic bile salt incubation, but not upon acid or bile salt alone. NF-kappa B pathway inhibition significantly decreased miRNA-143, -192, -194, COX2, and MUC2 expression. Additionally, miRNA-143, -145 and -194 expression was increased in reflux esophagitis biopsies compared with normal SQ, but no changes were found in miRNA-192 expression. Our findings suggest that upregulation of BE-specific miRNAs by acidic bile may be an early event in the transition of SQ to BE and that their expression is partly regulated by the NF-kappa B pathway.