Profiling of circulating microRNAs in patients with Barrett's esophagus and esophageal adenocarcinoma.

BACKGROUND: Circulating microRNAs (miRNAs) have been suggested as novel markers for various diseases. The goal of this pilot study was to identify circulating miRNAs differentially expressed comparing Barrett's esophagus (BE), esophageal adenocarcinoma (EAC), and controls. METHODS: MicroRNA expression profiling was performed by qPCR array using plasma from six controls and eight BE and eight EAC patients. Validation was performed by analyzing the expression of six selected miRNAs, by qRT-PCR in 115 plasma samples of controls, BE, and EAC patients. Diagnostic accuracy was evaluated by area under the curve (AUC) analysis. RESULTS: We identified three miRNAs that were elevated in EAC and four miRNAs that were elevated in BE. Further validation showed that miRNA-382-5p was significantly increased and miRNA-133a-3p significantly decreased in EAC. miRNA-194-5p and miRNA-451a were significantly increased and miRNA-136-5p significantly decreased in BE versus controls. A combination of three or more miRNAs was found to have a good diagnostic performance in discriminating BE from controls (AUC: 0.832), EAC from controls (AUC: 0.846), and BE from EAC (AUC: 0.797). CONCLUSION: Our data suggest that circulating miRNAs are differentially expressed in BE and EAC. The miRNAs identified may be used for future non-invasive screening of BE and EAC.

Prognostic factors for medium- and long-term survival of esophageal cancer patients in the Netherlands.

BACKGROUND AND OBJECTIVES: Medium- and long-term survival is low in esophageal cancer (EC) patients, which is thought to be due to tumor characteristics. Our aim was to determine both tumor- and non-tumor-related characteristics affecting survival in these patients. METHODS: Patients with primary EC between 1990 and 2008 in the southern part of the Netherlands were identified. Multivariable logistic regression was used to identify determinants of survival. RESULTS: In total, 703 patients with EC were included for the 1-year, 551 for the 3-year and 436 for the 5-year survival analysis. Poor 1-year survival was independently associated with chemoradiation (compared to surgery), positive lymph nodes (N1-stage) and 1 or ≥2 comorbidities. Adenocarcinoma (EAC) compared to squamous cell carcinoma was significantly associated with a better 1-year survival. Poor 3- and 5-year survival was associated with N1-stage and chemoradiation. Positive prognostic factors for 3- and 5-year survival were neoadjuvant therapy and female gender. CONCLUSION: Both tumor-related (negative lymph nodes and EAC histology) and non-tumor-related factors (surgery, neoadjuvant therapy, and female gender) are associated with a better survival of EC. Although it is not clear how histology and gender affect EC survival, knowledge of these factors may be relevant for clinical decision making.

Toll-like receptor 4 activation in Barrett's esophagus results in a strong increase in COX-2 expression.

BACKGROUND: Barrett's esophagus (BE) is known to progress to esophageal adenocarcinoma in a setting of chronic inflammation. Toll-like receptor (TLR) 4 has been linked to inflammation-associated carcinogenesis. We aimed to determine the expression and functional activity of TLR4 in the esophagus and whether TLR4 activation in BE could promote carcinogenesis by inducing COX-2 expression. METHODS: TLR4 expression in esophageal adenocarcinoma, BE, duodenum, reflux esophagitis and normal squamous esophagus biopsies was assessed using real-time PCR and validated by in situ hybridization and immunohistochemistry. Ex vivo cultures of BE, duodenum and normal squamous esophagus biopsies and a BE cell line (BAR-T) were stimulated with the TLR4 agonist lipopolysaccharide (LPS). To evaluate the effect of TLR4 activation, NF-κB activation, IL8 secretion and expression and COX-2 expression were determined. RESULTS: TLR4 expression was significantly increased in esophageal adenocarcinoma, BE, duodenum and reflux esophagitis compared to normal squamous esophagus. LPS stimulation resulted in NF-κB activation and a dose-dependent increase of IL8 secretion and mRNA expression. The induction of IL8 was more evident in BE compared to normal squamous esophagus. Upon LPS stimulation, COX-2 expression increased significantly in ex vivo cultured BE biopsies, which was observed in both epithelium and lamina propria cells. However, no effect was found in duodenum and normal squamous esophagus biopsies. CONCLUSION: TLR4 activation in BE results in a strong increase in COX-2 and may contribute to malignant transformation.

microRNA-145 in Barrett's oesophagus: regulating BMP4 signalling via GATA6.

OBJECTIVE: Barrett's oesophagus (BE) is a metaplastic condition of the distal oesophagus which predisposes to oesophageal adenocarcinoma (EAC). It has been suggested that microRNAs (miRNAs) are involved in the process of development of BE and EAC; however, few functional miRNA data are available. The aim of the study was to perform a tissue-specific miRNA profile and, based on this, to examine the function of miRNA-145 in the oesophagus. DESIGN: miRNA expression profiling using microarray analysis in EAC, BE and normal squamous epithelium of the oesophagus (SQ) was performed and validated using real-time PCR in samples from 15 patients and in situ hybridisation in samples from 10 patients. The proliferative effect of miRNA-145 precursor transfection in the SQ (HET-1A) and BE cell line (BAR-T) was measured. Downstream targets of miRNA-145 were determined by analysing mRNA and protein expression from miRNA-145 transfected cells. RESULTS: Three unique miRNA expression profiles were found in tissue from EAC, BE and SQ, which showed that miRNA-145 was upregulated in BE compared with EAC and SQ. Overexpression of miRNA-145 in HET-1A and BAR-T cells reduced cell proliferation and inhibited GATA6, BMP4 and SOX9 mRNA expression. Furthermore, altered BMP4 signalling was observed in vitro on miRNA-145 overexpression. These effects were blocked when cells were co-transfected with a miRNA-145 specific inhibitor. Additionally, BMP4 incubation of HET-1A cells altered miRNA-145 and GATA6 expression over time. CONCLUSION: These results imply that miRNA-145 indirectly targets BMP4 via GATA6 and is potentially involved in the development of BE.

The effect of socioeconomic status on staging and treatment decisions in esophageal cancer.

BACKGROUND: Optimal treatment choice for patients with esophageal cancer (EC) is complex and largely determined by tumor characteristics, comorbidity, and age. GOALS: This study describes the role of patient characteristics, among which is socioeconomic status (SES), in EC treatment. STUDY: Patients diagnosed with primary EC between 1990 and 2008 in the southern part of the Netherlands were identified using the Eindhoven Cancer Registry. Multivariable logistic and proportional hazard regression analyses were used to identify determinants of treatment and survival. RESULTS: We included 1914 patients, and 37% of them underwent intentionally curative treatment. Low-SES patients were diagnosed at older age (16% vs. 9%, age more than or equal to 80) and with more advanced tumor stages (13% vs. 10%, stage T4) than high-SES patients. Age less than 60 compared with 70 to 79 years [adjusted odds ratio, 4.51; 95% confidence interval (CI), 2.98-6.84] and high SES compared with low SES (adjusted odds ratio 1.59; 95% CI, 1.07-2.37) were independent predictors for curative treatment. Probability of death for high-SES patients undergoing palliative treatment was decreased compared with low-SES patients (hazard ratio, 0.84; 95% CI, 0.71-0.99). CONCLUSIONS: SES is an important factor in treatment choice of EC. As health care is equally accessible to the whole population in the Netherlands, this suggests that both patient-related and physician-related factors are involved in this phenomenon.