Abdel-Latif Mohamed Mahmoud M, Inoue Hiroyasu, Kelleher Dermot, Reynolds John V
Department of Surgery, Trinity Centre for Health Sciences, St. James's Hospital, Dublin 8, Ireland.
J Cancer Res Ther. 2016 Jan-Mar;12(1):364-73. doi: 10.4103/0973-1482.174525.
Gastroesophageal reflux disease is considered to be a major risk in the development of esophageal adenocarcinoma. Nuclear factor-kappa B (NF-κB) plays important roles in the regulation of several genes coding for cytokines, cell proliferation, and apoptosis. To understand the role of bile and acid in the causation of esophageal cancer, we have examined the effects of bile acids and acid on NF-κB activation in the esophageal epithelial cells OE33 and SKGT-4 qualitatively and quantitatively.
Analysis of NF-κB activation in esophageal epithelial cells in response to bile acids and acid was performed by electrophoretic mobility shift assay, Western blotting and the translocation NF-κB was assessed by high content analysis (HCA). Cyclooxygenase-2 (COX-2) promoter activity was assessed by transient transfection assays.
This study demonstrated that bile acids and acid activated NF-κB in a dose- and time-dependent manner. HCA analysis was an invaluable method in quantifying NF-κB translocation at the single cell population level following bile or acid treatment. Furthermore, deoxycholic acid (DCA) and acid-induced COX-2 promoter activity, and a mutation in the NF-κB and activator protein-1 (AP-1) binding sites remarkably reduced the reporter gene activity induced by DCA or acid.
Our data demonstrate that bile and acid induce NF-κB activation in esophageal cells qualitatively and quantitatively. The induction of COX-2 promoter activity by DCA and acid was mediated via NF-κB and AP-1 transcription. The activation of NF-κB signaling pathway in esophageal cells may contribute to the development of esophageal cancer, and, therefore, modulating of NF-κB pathway may uncover new therapeutic strategies.
胃食管反流病被认为是食管腺癌发生发展的主要危险因素。核因子-κB（NF-κB）在调控多种编码细胞因子、细胞增殖和凋亡的基因中发挥重要作用。为了解胆汁和酸在食管癌病因中的作用，我们定性和定量地研究了胆汁酸和酸对食管上皮细胞OE33和SKGT-4中NF-κB激活的影响。
通过电泳迁移率变动分析、蛋白质免疫印迹法分析食管上皮细胞对胆汁酸和酸反应时NF-κB的激活情况，并通过高内涵分析（HCA）评估NF-κB的易位。通过瞬时转染试验评估环氧化酶-2（COX-2）启动子活性。
本研究表明，胆汁酸和酸以剂量和时间依赖性方式激活NF-κB。HCA分析是定量胆汁或酸处理后单细胞群体水平上NF-κB易位的一种非常有价值的方法。此外，脱氧胆酸（DCA）和酸诱导COX-2启动子活性，而NF-κB和激活蛋白-1（AP-1）结合位点的突变显著降低了DCA或酸诱导的报告基因活性。
我们的数据表明，胆汁和酸在食管细胞中定性和定量地诱导NF-κB激活。DCA和酸对COX-2启动子活性的诱导是通过NF-κB和AP-1转录介导的。食管细胞中NF-κB信号通路的激活可能有助于食管癌的发展，因此，调节NF-κB通路可能揭示新的治疗策略。
