Novel metastatic models of esophageal adenocarcinoma derived from FLO-1 cells highlight the importance of E-cadherin in cancer metastasis

// David S. Liu 1, 2, 3 , Sanne J.M. Hoefnagel 4 , Oliver M. Fisher 5 , Kausilia K. Krishnadath 4, 6 , Karen G. Montgomery 1 , Rita A. Busuttil 1, 3, 7 , Andrew J. Colebatch 1 , Matthew Read 1, 2, 3 , Cuong P. Duong 1, 2, * , Wayne A. Phillips 1, 2, 3, 8, * , Nicholas J. Clemons 1, 3, * 1 Division of Cancer Research, Peter MacCallum Cancer Centre, Melbourne, Victoria, 3000, Australia 2 Division of Cancer Surgery, Peter MacCallum Cancer Centre, Melbourne, Victoria, 3000, Australia 3 Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, 3010, Australia 4 Laboratory for Experimental Oncology and Radiobiology, Center for Experimental and Molecular Medicine, Academic Medical Center, Amsterdam, 1105 AZ, The Netherlands 5 Gastroesophageal Cancer Program, St Vincent's Centre for Applied Medical Research, Darlinghurst, New South Wales, 2010, Australia 6 Department of Gastroenterology and Hepatology, Academic Medical Center, Amsterdam, 1105 AZ, The Netherlands 7 The University of Melbourne Department of Medicine, Royal Melbourne Hospital, Parkville, Victoria, 3010, Australia 8 University of Melbourne Department of Surgery, St Vincent's Hospital, Fitzroy, Victoria, 3065, Australia * Co-senior authors Correspondence to: Nicholas Clemons, email: nicholas.clemons@petermac.org Wayne Phillips, email: wayne.phillips@petermac.org Keywords: metastasis, esophageal cancer, CDH1, E-cadherin, animal models Received: September 06, 2016 Accepted: October 19, 2016 Published: November 16, 2016 ABSTRACT There is currently a paucity of preclinical models available to study the metastatic process in esophageal cancer. Here we report FLO-1, and its isogenic derivative FLO-1 LM , as two spontaneously metastatic cell line models of human esophageal adenocarcinoma. We show that FLO-1 has undergone epithelial-mesenchymal transition and metastasizes following subcutaneous injection in mice. FLO-1 LM , derived from a FLO-1 liver metastasis, has markedly enhanced proliferative, clonogenic, anti-apoptotic, invasive, immune-tolerant and metastatic potential. Genome-wide RNAseq profiling revealed a significant enrichment of metastasis-related pathways in FLO-1 LM cells. Moreover, CDH1 , which encodes the adhesion molecule E-cadherin, was the most significantly downregulated gene in FLO-1 LM compared to FLO-1. Consistent with this, repression of E-cadherin expression in FLO-1 cells resulted in increased metastatic activity. Importantly, reduced E-cadherin expression is commonly reported in esophageal adenocarcinoma and independently predicts poor patient survival. Collectively, these findings highlight the biological importance of E-cadherin activity in the pathogenesis of metastatic esophageal adenocarcinoma and validate the utility of FLO-1 parental and FLO-1 LM cells as preclinical models of metastasis in this disease.