The miR-361–3p regulates RelA and CDX2 mRNA expression; and miR-212–3p regulates COX-2 mRNA expression. miRNAs may contribute to the inflammation of lower esophagus with H. pylori infection, and may be involved in the development of Barrett’s Esophagus and esophageal adenocarcinoma. Key Word(s): 1. MicroRNAs; 2. COX-2; 3. CDX2; 4. Helicobacter pylori; Presenting Author: GUI-GEN TENG Additional Authors: WEI-HONG WANG, YUN DAI, SHU-JUN WANG, YUN-XIANG CHU, JIANG LI Corresponding Author: WEI-HONG WANG Affiliations: Peking University First Hospital Objective: Barrett’s esophagus (BE) is recognized as a complication of chronic gastroesophageal acid
and bile reflux, and also considered to be a precancerous state in the esophagus and may progress to esophageal adenocarcinoma (EA). H. pylori has been found to colonize the Barrett epithelium of lower esophagus. However, GS 1101 its role in the development of Barrett’s buy MK-2206 esophagus and esophageal adenocarcinoma is unclear. Here, we explored the effects of acidic deoxycholic acid and H. pylori on esophageal cell lines in vitro, with a particular focus on whether NF-kB is involved in this event. Methods: H. pylori 26695 and its cagA mutant strain were cocultured
with two esophageal cell lines (HET-1A, OE33) with or without acidic deoxycholic acid (DCA) in vitro. Cell proliferation was tested by CCK-8 assay. Apoptosis was determined by Flow Cytometry. COX-2 and CDX2 were assessed by real-time PCR and Western blot. MUC2 was determined by qPCR and immunocytochemistry. MCE公司 NF-kB phosphorylation and
DNA-binding activity were determined by Western blot and EMSA. NF-kB transcriptional activity was identified by luciferase reporter assay. The downstream genes of NF-kB, such as IL-8 were assessed by ELISA and qPCR. Results: DCA, live H. pylori and HPE (H. pylori extract) reduced proliferation and promoted apoptosis of esophageal cells. DCA, live H. pylori and HPE up-regulate the expression of COX-2, CDX2 and MUC2 in both esophageal cell lines. However, cagA mutant strain and its extract did not induce the expression of CDX2 and MUC2. NF-kB activity was induced by DCA, live H. pylori and HPE. Treatment with DCA in the presence of either live H. pylori or HPE further augmented the NF-kB phosphorylation, its DNA-binding and the transcriptional activity; and subsequently increased the expression of COX-2, CDX2, MUC2 and IL-8 as compared to DCA treatment alone. The results suggested a synergistic effect between DCA and H. pylori in esophageal cells. Both siRNA P65 and PDTC significantly inhibited NF-kB activity, and influenced the downstream genes expression in esophageal cell lines with H. pylori infection. Conclusion: The present study reveals that both H. pylori and DCA up-regulate the expression of COX-2, CDX2, MUC2 and IL-8 in esophageal cells by inducing the activation of NF-kB. These results indicate that H.