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

Abstract

Background:Barrett’s oesophagus is a pre-malignant condition at gastroesophageal junction in which normal squamous epithelium is replaced by columnar shape epithelium, which predisposes oesophageal adenocarcinoma. It is known that Barrett’s oesophagus evolves as a consequence of chronic gastro-oesophageal reflux disease. Although progression of Barrett’s oesophagus to adenocarcinoma is still unclear, increasing incidence of oesophageal cancer and mortality worldwide make its study necessary. Several investigations have been made on the aetiology of oesophageal cancer. Most of them assessed genetical or environmental factors. However, potential role of bacteria in the development of oesophageal adenocarcinoma as a new environmental factor has not been addressed. Previous study on Barrett’s disease detected presence of Campylobacter concisus as a new emerging pathogen on Barrett’s and oesophageal cancer samples compared with healthy individuals. This indicates that this organism might involve in the progression of Barrett’s to oesophageal adenocarcinoma.Objectives:This study aimed to determine the effects of C. concisus on expression of three biomarkers including interleukin-18 (IL-18), tumour necrosis factor-α (TNF-α) and tumour suppressor gene (p53) in three Barrett's cell lines.Materials and Methods:Quantitative real-time PCR assays were developed to measure expression of pro-inflammatory mediators (IL-18 and TNF-α) and gene expression of p53 in Barrett's cell lines in co-culture with C. concisus.Results:The mentioned organism was able to modulate considerably expression of p53, TNF-α and IL-18 in a time-dependent manner.Conclusions:The results showed that microorganism influences expression of carcinogenesis biomarker and cytokines in cell line models and possibility promotes oesophageal adenocarcinoma.

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

“…This study planned to determine the consequences of probiotics on the expression of key biomarkers that contribute to the progression of BO to OA. These molecular markers were identified as being modulated by the effect of C. concisus in our previous study that identified C. concisus as a putative pathogen in the transformation of BO to OA (Mozaffari namin et al, 2015). This study indicates evidence on the inhibitory effect of probiotics to prevent progression of BO to OA in an in vitro model which might help new exploration into OA treatment.…”

“…B. longum and Lactobacillus acidophilus strains were selected as probiotics and the strain Campylobacter concisus ATCC13826 was selected as a pathogen and was used previously by Mozaffari namin et al (2015). The identity of the probiotic strains was verified by 16S rRNA gene sequence analyses (DNA analysis lab at Ninewells Hospital) and a search using BLAST confirmed the strains to be B. longum KACC 91563 and L. acidophilus NCFM.…”

“…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 (Campylobacter concisus) may initiate the progression of BO (Mozaffari namin et al, 2015) Despite the availability of several diagnostic techniques and treatment protocols, there are conflicting results on screening and surveillance of BO and OA survival (Lim & Fitzgerald, 2013;Sharma et al, 2013;Thrift et al, 2014). The use of probiotics has been established in the prevention of carcinogenic diseases.…”

“…Pre-neoplastic BO cell lines including CP-A (nondysplastic metaplasia) and CP-D (high-grade dysplastic) were selected for co-culturing with the probiotics as described previously (Mozaffari namin et al, 2015). In brief, Barrett's cell lines were maintained in keratinocyte serum-free medium (Invitrogen) with supplements.…”

“…The mRNA gene expression of selected biomarkers was detected following co-culture with probiotics and assessed against the GAPDH housekeeping gene by extraction of mRNA, synthesis of cDNA using primer sets described in Table 1 and quantified by real-time quantitative PCR (qPCR) as described previously (Mozaffari namin et al, 2015). DNA standards for quantification of DNA were prepared and qPCR was then performed by using an iCycler real-time PCR Detection System (Bio-Rad Laboratories) by iQ SYBR Green Super mix (Bio-Rad) with appropriate annealing temperature (Table 1).…”