Ephrin B3 interacts with multiple EphA receptors and drives migration and invasion in non-small cell lung cancer

// Ghazal Efazat 1, * , Metka Novak 1, * , Vitaliy O. Kaminskyy 2 , Luigi De Petris 1 , Lena Kanter 1 , Therese Juntti 1 , Per Bergman 3 , Boris Zhivotovsky 2 , Rolf Lewensohn 1 , Petra Hååg 1 , Kristina Viktorsson 1 1 Karolinska Biomics Center, Department of Oncology-Pathology, Karolinska Institutet, SE-171 76 Stockholm, Sweden 2 Institute of Environmental Medicine, Division of Toxicology, Karolinska Institutet, SE-171 77 Stockholm, Sweden 3 Department of Molecular Medicine and Surgery (MMK), Thoracic Surgery, Karolinska Institutet, SE-171 76 Stockholm, Sweden * These authors have contributed equally to this work Correspondence to: Kristina Viktorsson, email: Kristina.Viktorsson@ki.se Petra Hååg, email: Petra.Haag@ki.se Keywords: Ephrin B3, EphA2, NSCLC, migration, invasion Received: September 17, 2015     Accepted: July 16, 2016     Published: August 11, 2016 ABSTRACT Ephrin receptors (Ephs) are reported to control metastatic signaling of non-small cell lung cancer (NSCLC) and other tumors. Here we show for the first time that blocking expression of the Eph ligand Ephrin B3 inhibits NSCLC cell migration and invasion. We demonstrate that Ephrin B3 directly binds the EphAs EphA2, EphA3, EphA4, and EphA5. EphA2 Ser897 was previously shown to drive migration propensity of tumor cells and our study reveals that EphA2 stays phosphorylated on Ser897 in the Ephrin B3/EphA2 complex in NSCLC cells of different histology. Moreover, we report that within such Ephrin B3/EphA2 complex both Akt Ser 129 and p38MAPK are found indicating a potential to drive migration/proliferation. We also found the EMT marker E-cadherin expression to be maintained or increased upon Ephrin B3 blockade in NSCLC cells. Expression of Ephrin B3 was furthermore analyzed in a cohort of NSCLC stage IA-IB cases (n=200) alongside EphA2 and Ephrin A1. We found that Ephrin B3 was concomitantly expressed with EphA2 and Ephrin A1 with higher Ephrin B3 levels found in non-squamous than in squamous tumors, whereas EphA2 was higher expressed in well-differentiated than in low-differentiated tumors. In the entire NSCLC cohort, Ephrin B3 expression was not linked to patient survival, whereas a high EphA2 expression was associated with improved survival (p=0.03). In conclusion, we show that blocking Ephrin B3 expression inhibits NSCLC proliferation-, migration- and invasion capacity which calls for further studies on interference with Ephrin B3 as a possible therapeutic avenue in this tumor malignancy.