In vitro studies Pritelivir nmr have shown NET1 expression to drive invasion in gastric adenocarcinoma [12]. Separately it has also been shown to be functionally important in epithelial mesenchymal transition in retinal epithelial cells [13], keratinocytes [14] and during gastrulation [15]. NET1 has previously been shown to be differentially expressed and functionally important in mediating cancer cell invasion in gastric cancer [12, 16] and in

squamous cell skin cancer (17). It has also been shown to be prominent in a number of other cancers [17–21] and to be a marker of poor prognosis in many of these (Table 1). Our group have previously shown NET1 to be of functional importance in breast and gastric cancer [4, 12, 16, 22]. Recognising the mounting cellular and molecular evidence for a role for NET1 in mediating gastrointestinal (GI) cancers and coupled with the phenotypic similarities recognised in the pathogenesis of gastric and oesophageal adenocarcinomas [1], we sought to investigate and fully characterise the bioactivity of NET 1 in oesophageal cancer. Table 1 A summary of current data on NET1 in other human cancers Cancer type Role of NET1 Reference Gastric adenocarcinoma Invasion via RhoA Leyden et al. [12] Murray et al. [4] Breast cancer Predicts late stage and poor prognosis

Gilcrease Doramapimod et al. [18] Mediates morphine-induced cell migration in vitro Ecimovic et al. [22] Glioma Marker of invasion and aggressive disease. Poorer survival in NET1 positive Tu et al. [20] Hepatocellular carcinoma

Correlates with higher histological grade Chen et al. [17] Cervical carcinoma Highly expressed in cervical epithelial neoplasia and in carcinoma Wollscheid et al. [21] Methods Cell culture Our in vitro oesophageal cell line model consisted of six cell lines: Het1a an SV40 immortalised normal oesophageal cell line derived from a 25 year old male; two Barrett’s cell lines QhTERT and GihTERT previously established by hTERT immortalisation (American Type Culture Collection, Virginia, USA) that represent non-dysplastic and high grade dysplastic Barrett’s epithelium respectively; and three Barrett’s related oesophageal adenocarcinoma cell lines – OE33, OE19 Obatoclax Mesylate (GX15-070) and JH-EsoAd1. OE33 was established from an adenocarcinoma of the lower esophagus of a 73-year-old female patient and is pathological stage IIA and poorly differentiated. OE19 is a pathological stage III moderately differentiated adenocarcinoma of gastric cardia/oesophageal gastric junction in a 72-year-old male patient. JH-EsoAD1 is from a patient with Barrett’s associated adenocarcinoma [23]. AGS is a gastric cancer cell line from a 54 year old female and represents a moderate to poorly differentiated adenocarcinoma. SW480 is from a locally invasive (Duke’s stage B) colon adenocarcinoma.