Gene expression changes associated with Barrett's esophagus and Barrett's-associated adenocarcinoma cell lines after acid or bile salt exposure

Abstract Cardiac-type epithelium has been proposed as an intermediate stage between normal squamous epithelium and intestinal metaplasia in the development of Barrett’s esophagus. Deregulation of certain miRNAs and their effects on CDX2 expression might contribute to the intestinalization process of cardiac-type epithelium. The aim of this study was to identify miRNAs differentially expressed between CDX2 positive and negative glands of Barrett’s esophagus and to examine the function of specific miRNAs on the regulation of CDX2. Methods miRNA expression profiling using OpenArrayTM analysis in microdissected cardiac-type glands with and without fully CDX2 expression was performed in biopsies from patients who developed cardiac-type epithelium in the remnant esophagus after esophagectomy. Data were validated using real-time PCR in esophageal adenocarcinoma cell lines and in situ and real-time PCR miRNA/CDX2/MUC2 co-expression analysis in cardiac-type mucosa samples. The effect of miR-24-3p precursor transfection on CDX2 expression was assessed in the esophageal adenocarcinoma cell lines FLO-1 and KYAE-1. Results CDX2 positive glands were characterized by an unique miRNA profile with a significant downregulation of miR-24-3p, miR-520e-3p, miR-548a-1, miR-597-5p, miR-133a-3p, miR-30a-5p, miR-638, miR-625-3p, miR-1255b-1, miR-1260a and upregulation of miR-590 (Figure 1A). miRNA-24-3p was identified as potential regulator of CDX2 gene expression in three bioinformatics algorithms, and this was confirmed in esophageal adenocarcinoma cell lines (Figure 1C). Furthermore, miR-24-3p expression negatively correlates with CDX2 in cardiac-type mucosa samples with different stages of mucosal intestinalization (Figure 1B). Conclusion These results imply that miRNA-24-3p directly targets CDX2, and downregulation of miRNA-24-3p is associated with the acquisition of an intestinal phenotype in cardiac-type epithelium.

Download Full-text

In Esophageal Squamous Cell Lines Derived From GERD Patients With and Without Barrett's Esophagus, There Are Differences in NF-κB Activation After Acid and Bile Salt Exposure

Gastroenterology ◽

10.1016/s0016-5085(11)60307-7 ◽

2011 ◽

Vol 140 (5) ◽

pp. S-76 ◽

Cited By ~ 1

Author(s):

Xiaofang Huo ◽

Xi Zhang ◽

Chunhua Yu ◽

Edaire Cheng ◽

David H. Wang ◽

...

Keyword(s):

Bile Salt ◽

Barrett’S Esophagus ◽

Cell Lines ◽

Squamous Cell ◽

Barrett's Esophagus

Download Full-text

miR-24-3p regulates CDX2 during intestinalization of cardiac-type epithelium in a human model of Barrett’s esophagus

Diseases of the Esophagus ◽

10.1093/dote/doab005 ◽

2021 ◽

Author(s):

Gabriel Gil-Gómez ◽

Matteo Fassan ◽

Lara Nonell ◽

Marta Garrido ◽

Marta Climent ◽

...

Keyword(s):

Barrett’S Esophagus ◽

Esophageal Adenocarcinoma ◽

Cell Lines ◽

Messenger Rna ◽

Barrett's Esophagus ◽

Human Model ◽

Adenocarcinoma Cell ◽

Intestinal Phenotype ◽

Cdx2 Expression

Summary Background Cardiac-type epithelium has been proposed as the precursor of intestinal metaplasia in the development of Barrett’s esophagus. Dysregulation of microRNAs (miRNAs) and their effects on CDX2 expression may contribute to intestinalization of cardiac-type epithelium. The aim of this study was to examine the possible effect of specific miRNAs on the regulation of CDX2 in a human model of Barrett’s esophagus. Methods Microdissection of cardiac-type glands was performed in biopsy samples from patients who underwent esophagectomy and developed cardiac-type epithelium in the remnant esophagus. OpenArray™ analysis was used to compare the miRNAs profiling of cardiac-type glands with negative or fully positive CDX2 expression. CDX2 was validated as a miR-24 messenger RNA target by the study of CDX2 expression upon transfection of miRNA mimics and inhibitors in esophageal adenocarcinoma cell lines. The CDX2/miR-24 regulation was finally validated by in situ miRNA/CDX2/MUC2 co-expression analysis in cardiac-type mucosa samples of Barrett’s esophagus. Results CDX2 positive glands were characterized by a unique miRNA profile with a significant downregulation of miR-24-3p, miR-30a-5p, miR-133a-3p, miR-520e-3p, miR-548a-1, miR-597-5p, miR-625-3p, miR-638, miR-1255b-1, and miR-1260a, as well as upregulation of miR-590-5p. miRNA-24-3p was identified as potential regulator of CDX2 gene expression in three databases and confirmed in esophageal adenocarcinoma cell lines. Furthermore, miR-24-3p expression showed a negative correlation with the expression of CDX2 in cardiac-type mucosa samples with different stages of mucosal intestinalization. Conclusion These results showed that miRNA-24-3p regulates CDX2 expression, and the downregulation of miRNA-24-3p was associated with the acquisition of the intestinal phenotype in esophageal cardiac-type epithelium.

Download Full-text

Gene expression profile comparison of Barrett's esophagus epithelial cell cultures and biopsies

Diseases of the Esophagus ◽

10.1111/j.1442-2050.2008.00810.x ◽

2008 ◽

Vol 21 (7) ◽

pp. 628-633 ◽

Cited By ~ 2

Author(s):

J. W. P. M. van Baal ◽

A. M. Rygiel ◽

F. Milano ◽

M. Anderson ◽

J. J. G. H. M. Bergman ◽

...

Keyword(s):

Gene Expression ◽

Epithelial Cell ◽

Barrett’S Esophagus ◽

Gene Expression Profile ◽

Cell Cultures ◽

Expression Profile ◽

Barrett's Esophagus ◽

Epithelial Cell Cultures

Download Full-text

Identification of miRNAs and genes for predicting Barrett’s esophagus progressing to esophageal adenocarcinoma using miRNA-mRNA integrated analysis

PLoS ONE ◽

10.1371/journal.pone.0260353 ◽

2021 ◽

Vol 16 (11) ◽

pp. e0260353

Author(s):

Chengjiao Yao ◽

Yilin Li ◽

Lihong Luo ◽

Qin Xiong ◽

Xiaowu Zhong ◽

...

Keyword(s):

Gene Expression ◽

Barrett’S Esophagus ◽

Esophageal Adenocarcinoma ◽

Barrett's Esophagus ◽

Trp Channels ◽

Functional Enrichment ◽

Integrated Analysis ◽

Epstein Barr Virus Infection ◽

Tissue Samples ◽

Microarray Datasets

Barrett’s esophagus (BE) is defined as any metaplastic columnar epithelium in the distal esophagus, which predisposes to esophageal adenocarcinoma (EAC). Yet, the mechanism through which BE develops to EAC still remain unclear. Moreover, the miRNA-mRNA regulatory network in distinguishing BE from EAC still remains poorly understood. To identify differentially expressed miRNAs (DEMs) and genes (DEGs) between EAC and BE from tissue samples, gene expression microarray datasets GSE13898, GSE26886, GSE1420 and miRNA microarray datasets GSE16456, GSE20099 were downloaded from Gene Expression Omnibus (GEO) database. GEO2R was used to screen the DEMs and DEGs. Pathway and functional enrichment analysis were performed by DAVID database. The protein–protein interaction (PPI) network was constructed by STRING and been visualized by Cytoscape software. Finnal, survival analysis was performed basing TCGA database. A total of 21 DEMs were identified. The enriched functions and pathways analysis inclued Epstein-Barr virus infection, herpesvirus infection and TRP channels. GART, TNFSF11, GTSE1, NEK2, ICAM1, PSMD12, CTNNB1, CDH1, PSEN1, IL1B, CTNND1, JAG1, CDH17, ITCH, CALM1 and ITGA6 were considered as the hub-genes. Hsa-miR-143 and hsa-miR-133b were the highest connectivity target gene. JAG1 was predicted as the largest number of target miRNAs. The expression of hsa-miR-181d, hsa-miR-185, hsa-miR-15b, hsa-miR-214 and hsa-miR-496 was significantly different between normal tissue and EAC. CDH1, GART, GTSE1, NEK2 and hsa-miR-496, hsa-miR-214, hsa-miR-15b were found to be correlated with survival.

Download Full-text

Global Changes in Gene Expression of Barrett's Esophagus Compared to Normal Squamous Esophagus and Gastric Cardia Tissues

PLoS ONE ◽

10.1371/journal.pone.0093219 ◽

2014 ◽

Vol 9 (4) ◽

pp. e93219 ◽

Cited By ~ 15

Author(s):

Paula L. Hyland ◽

Nan Hu ◽

Melissa Rotunno ◽

Hua Su ◽

Chaoyu Wang ◽

...

Keyword(s):

Gene Expression ◽

Barrett’S Esophagus ◽

Barrett's Esophagus ◽

Gastric Cardia ◽

Global Changes

Download Full-text

Gene Expression Profiling Reveals Stromal Genes Expressed in Common Between Barrett’s Esophagus and Adenocarcinoma

Gastroenterology ◽

10.1053/j.gastro.2006.04.026 ◽

2006 ◽

Vol 131 (3) ◽

pp. 925-933 ◽

Cited By ~ 96

Author(s):

Ying Hao ◽

George Triadafilopoulos ◽

Peyman Sahbaie ◽

Harvey S. Young ◽

M. Bishr Omary ◽

...

Keyword(s):

Gene Expression ◽

Gene Expression Profiling ◽

Barrett’S Esophagus ◽

Expression Profiling ◽

Barrett's Esophagus

Download Full-text

Bile salts induce or blunt cell proliferation in Barrett's esophagus in an acid-dependent fashion

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.2000.278.6.g1000 ◽

2000 ◽

Vol 278 (6) ◽

pp. G1000-G1009 ◽

Cited By ~ 55

Author(s):

Baljeet S. Kaur ◽

Rodica Ouatu-Lascar ◽

M. Bishr Omary ◽

George Triadafilopoulos

Keyword(s):

Cell Proliferation ◽

Bile Salt ◽

Barrett’S Esophagus ◽

Barrett's Esophagus ◽

Bile Salts ◽

Bile Reflux ◽

Salt Mixture ◽

Complete Obliteration ◽

Protein Kinase C Inhibitor ◽

Incorporation Assay

Barrett's esophagus (BE) results from acid and bile reflux and predisposes to cancer. We investigated the effect of bile salts, with or without acid, on cell proliferation in BE and assessed mechanism(s) involved. To mimic physiological conditions, biopsies of esophagus, BE, and duodenum were exposed to a bile salt mixture, either continuously or as a 1-h pulse, and were compared with control media without bile salts (pH 7.4) for ≤24 h. Similar experiments were also performed with acidified media (pH 3.5) combined with the bile salt mixture as a 1-h pulse. Cell proliferation was assessed by a [3H]thymidine incorporation assay with or without bisindolylmaleimide (BIM), a selective protein kinase C inhibitor. Bile salt pulses enhanced cell proliferation in BE without affecting cell proliferation in esophageal or duodenal epithelia. In the presence of BIM, there was complete obliteration of the bile salt-induced BE hyperproliferation. In contrast, 1-h pulses of bile salts in combination with acid significantly inhibited proliferation in BE but had no effect on esophagus or duodenum. We conclude that in BE explants, brief exposure to bile salts, in the absence of acid, increases proliferation, whereas exposure to a combination of bile salts and acid together inhibits proliferation.

Download Full-text

In Barrett’s epithelial cells, weakly acidic bile salt solutions cause oxidative DNA damage with response and repair mediated by p38

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00329.2019 ◽

2020 ◽

Vol 318 (3) ◽

pp. G464-G478

Author(s):

Xiaofang Huo ◽

Kerry B. Dunbar ◽

Xi Zhang ◽

Qiuyang Zhang ◽

Stuart Jon Spechler ◽

...

Keyword(s):

Dna Damage ◽

Bile Salt ◽

Barrett’S Esophagus ◽

Esophageal Adenocarcinoma ◽

Dna Damage Response ◽

Barrett's Esophagus ◽

Oxidative Dna Damage ◽

Damage Response ◽

Barrett's Metaplasia ◽

Barrett’S Metaplasia

The frequency of esophageal adenocarcinoma is rising despite widespread use of proton pump inhibitors (PPIs), which heal reflux esophagitis but do not prevent reflux of weakly acidic gastric juice and bile in Barrett’s esophagus patients. We aimed to determine if weakly acidic (pH 5.5) bile salt medium (WABM) causes DNA damage in Barrett’s cells. Because p53 is inactivated frequently in Barrett’s esophagus and p38 can assume p53 functions, we explored p38’s role in DNA damage response and repair. We exposed Barrett’s cells with or without p53 knockdown to WABM, and evaluated DNA damage, its response and repair, and whether these effects are p38 dependent. We also measured phospho-p38 in biopsies of Barrett’s metaplasia exposed to deoxycholic acid (DCA). WABM caused phospho-H2AX increases that were blocked by a reactive oxygen species (ROS) scavenger. WABM increased phospho-p38 and reduced bromodeoxyuridine incorporation (an index of S phase entry). Repair of WABM-induced DNA damage proceeded through p38-mediated base excision repair (BER) associated with reduction-oxidation factor 1-apurinic/apyrimidinic endonuclease I (Ref-1/APE1). Cells treated with WABM supplemented with ursodeoxycholic acid (UDCA) exhibited enhanced p38-mediated responses to DNA damage. All of these effects were observed in p53-intact and p53-deficient Barrett’s cells. In patients, esophageal DCA perfusion significantly increased phospho-p38 in Barrett’s metaplasia. WABM exposure generates ROS, causing oxidative DNA damage in Barrett’s cells, a mechanism possibly underlying the rising frequency of esophageal adenocarcinoma despite PPI usage. p38 plays a central role in oxidative DNA damage response and Ref-1/APE1-associated BER, suggesting potential chemopreventive roles for agents like UDCA that increase p38 activity in Barrett’s esophagus. NEW & NOTEWORTHY We found that weakly acidic bile salt solutions, with compositions similar to the refluxed gastric juice of gastroesophageal reflux disease patients on proton pump inhibitors, cause oxidative DNA damage in Barrett’s metaplasia that could contribute to the development of esophageal adenocarcinoma. We also have elucidated a critical role for p38 in Barrett’s metaplasia in its response to and repair of oxidative DNA damage, suggesting a potential chemopreventive role for agents like ursodeoxycholic acid that increase p38 activity in Barrett’s esophagus.

Download Full-text

PS02.057: INDUCTION OF APOPTOSIS IN ESOPHAGEAL ADENOCARCINOMA CELLS BY CURCUMIN

Diseases of the Esophagus ◽

10.1093/dote/doy089.ps02.057 ◽

2018 ◽

Vol 31 (Supplement_1) ◽

pp. 136-137

Author(s):

René Thieme ◽

Florian Michiels ◽

Luisa Maurer ◽

Albrecht Hoffmeister ◽

Uwe Eichfeld ◽

...

Keyword(s):

Barrett’S Esophagus ◽

Cell Lines ◽

Stable Disease ◽

Barrett's Esophagus ◽

Conflicts Of Interest ◽

Curcuma Longa ◽

Inflammatory Microenvironment ◽

Induction Of Apoptosis ◽

Anti Inflammatory ◽

Proliferation Assays

Abstract Background Curcumin naturally occurring in curry powder from Curcuma longa is an anti-inflammatory and anti-proliferative agent. It represses NFκB activation and induces apoptosis in cancer cells. Because, inflammation favours the onset of an intestinal metaplasia in esophageal squamous epithelial cells and esophageal carcinoma cells, a reduction of such processes may contribute to establish a stable disease and/or sensitization for chemotherapeutic approaches. Methods Curcumin receptivity was investigated in metaplastic (CP-A), dysplastic (CP-B), adenocarcinoma (OE19, OE33), and esophageal fibroblast (FFE3) cell lines, which were treated with 10 or 25 μM Curcumin for 48h or 72h. Response to Curcumin was measured by proliferation assays as well as by induction of apoptosis and Akt activation by western blot analyses. Results The EAC cell lines OE33 and OE19 show a decrease of proliferation with raising curcumin concentration with an IC50 of 9.6 and 14.8 μM, respectively. While the metaplastic and dysplastic cell lines showed a comparable IC50 of 7.7 and 11.6 μM to the EAC cell lines, the FEF3 cell showed a higher IC50 of approx. 20μM. The phosphorylation of Akt was decreased and apoptosis was induced, showing cleaved PARP, when treated with 25μM curcumin after 48h and 72h. Conclusion Metaplastic, dysplastic and EAC cells show a higher receptivity to curcumin than esophageal fibroblasts cells. This constrains the NFκB activation and its contribution in the manifestation of the Barrett's esophagus. The usage of curcumin to generate an anti-inflammatory microenvironment will potentially help to develop a stable disease or to reverse the development of the Barrett's esophagus. Disclosure All authors have declared no conflicts of interest.

Download Full-text