Effect of probiotics on the expression of Barrett’s oesophagus biomarkers

Abstract

Barrett's oesophagus (BO) is a complicated condition at the gastroesophageal junction in which normal squamous epithelium is changed to columnar and leads to oesophageal adenocarcinoma (OA). In the past decades, the prevalence of Barrett's disease and mortality rate of adenocarcinoma has significantly increased throughout the word. Data has shown that molecular pathogenesis of disease has not been clearly identified. However, a wide-range and successful administration of probiotics in cancer and gastrointestinal diseases has lead to the investigation into the possible inhibitory role of probiotics in oesophageal cancer. This study was conducted to evaluate the inhibitory effect of probiotics on the expression of biomarkers in an in vitro model. Two different Barrett's oesophageal cell lines were selected to co-culture with B. longum and Lactobacillus acidophilus to measure expression of IL-18, TNFa, p53 (tumour suppressor gene), cyclooxygenase 2 and CDX1 (caudal type homeobox 1) genes. In addition, two different aspects of probiotic administration, therapeutic and prophylactic test were also examined. Results showed that micro-organisms could inhibit expression of biomarkers and therapeutic culture conditions were more effective than prophylactic tests. The results obtained suggest that it is possible to incorporate the administration of probiotics in BO and OA prevention. INTRODUCTIONBarrett's oesophagus (BO) is well recognized as a precursor of oesophageal adenocarcinoma (OA), which is usually caused by gastroesophageal reflux disease (GORD). In BO, normal squamous epithelium is replaced by columnar intestinal epithelium because of GORD effects. It is known that progression of BO to OA includes sequence of metaplasia-dysplasia-adenocarcinoma, which is created by the effect of various factors and multiple pathways including environmental and genetic factors. The incidences of both Barrett's and oesophageal cancer have risen in the world particularly in the Western world and the UK by variation between 1/52 and 1/694, respectively (HvidJensen et al., 2011;Werner & Laßmann, 2012;Zimmerman, 2014). Subsequently, different studies have evaluated a range of putative factors that might support risk of progression of BO to OA. Current evidence suggests that progression of Barrett's disease is associated with the changes in bacterial population in GORD, BO and OA patients (Blackett et al., 2013). It has been suggested that Gram-negative bacteria (Lim & Fitzgerald, 2013;Sharma et al., 2013;Thrift et al., 2014). The use of probiotics has been established in the prevention of carcinogenic diseases. Probiotics are described as live micro-organisms applied in adequate amounts for beneficial use in order to strengthen the ordinary defence system and protect the gastrointestinal epithelium against pathogenic bacteria. Probiotics have been used successfully in cases of infant diarrhoea, food allergies, ulcerative colitis, colonic cancer, Crohn's disease and inflammatory bowel disease (Furrie et al., 2005;Steed et al., 2010;Ri...

The Effect of Campylobacter concisus on Expression of IL-18, TNF-α and p53 in Barrett’s Cell Lines

“…Bacteria were then centrifuged (2500 × g, 25 minutes), washed once by PBS, counted in haemocytometer and resuspended to concentration of 3 × 107 ml −1 . S. salivarius was grown aerobically in WC broth at 37°C for use as a negative control, as part of the normal human oesophageal microbiota ( 16 ).…”

“…The Barrett’s associated adenocarcinoma cell line FLO-1 (immortalised Barrett's oesophageal epithelial cells) and two Barrett’s cell lines (CPA (non-dysplastic metaplasia) and CPD (high-grade dysplastic metaplasia)) were used to challenge with organism as described by Mozaffari Namin et al ( 16 ). Briefly, FLO-1 was maintained in Dulbecco’s modified eagle medium (DMEM) (Sigma-Aldrich, Gillingham, UK) with 10% foetal calf serum (FCS) (Gibco-BRL life technologies, UK) and 1% penicillin/ treptomycin (Gibco-BRL, UK).…”

“…Gene expression profile of biomarkers were determined by relative target gene quantities from threshold cycles (C t ) normalised against the housekeeping gene GAPDH in FLO-1and BO cell lines through the mRNA extraction using the RNA easy kit (Qiagen, Crawley, UK), cDNA synthesis using the Quick reverse transcription system (Promega) and quantification by real-time PCR (qPCR) with primer sets after stimulating by C. concisus as described previously by Mozaffari Namin et al ( 16 ). Briefly, products of correct size and sequence were purified using the Qiaquick PCR purification kit and ligated into a vector using the pGEM-T easy vector system I (Promega).…”

Changes in the distal esophageal microbiota in Chinese patients with reflux esophagitis

“…Another in vitro model study showed that the expression of tumor necrosis factor‐α and p53 was significantly reduced when two different Barrett's esophagus cell lines were co‐cultured with Bifidobacterium longum , and the inhibitory effect of probiotics might play a role in preventing the progression of Barrett's esophagus to adenocarcinoma . Our current preliminary investigation (unpublished) showed that the composition of the esophageal mucosal flora of the patients with esophageal squamous cell carcinoma ( n = 9) were significantly changed compared with controls ( n = 17).…”

“…22 Another in vitro model study showed that the expression of tumor necrosis factor-α and p53 was significantly reduced when two different Barrett's esophagus cell lines were co-cultured with Bifidobacterium longum, and the inhibitory effect of probiotics might play a role in preventing the progression of Barrett's esophagus to adenocarcinoma. 23 Our current preliminary investigation (unpublished) showed We found that esophageal mucosa contains many bacteria, and four strains of bacteria were significantly lower in patients with RE (Prevotella, Helicobacter, and Moraxella were reduced at the genus level, and Bacteroidetes was reduced at the phylum level). There was no significant difference between the two groups with respect to other genera.…”