The molecular and cellular impact of EPLIN(Epithelial Protein Lost in Neoplasm) on the healingof human wounds

EPLIN (Epithelial Protein Lost In Neoplasm) is a cytoskeletal associated protein whose expression is often reduced in cancer cells. It may function as a tumour suppressor through its effects on cancer cell migration and invasion. To date, its role in wound healing has not been elucidated. We examine the impact of EPLIN on keratinocyte migration and its implications in wound healing. A mammalian expression construct containing the full EPLIN coding sequence was used to overexpress EPLIN in human keratinocyte cell (HaCaT). Following overexpression verification, the impact of EPLIN on HaCaT cell migration was assessed using a conventional scratch wounding assay and an electric cell-substrate impedance sensing (ECIS) system-based assay. Protein expression was examined using western blot, ICC and IFC analysis. Transfection of HaCaT cells with the EPLIN expression construct successfully resulted in enhanced HaCaT EPLIN expression. Enhanced EPLIN levels were seen to negatively impact on cell migration as determined by both the scratch wound assay and the ECIS model system with migration rates of HaCaT cells overexpressing EPLIN being substantially less than the control HaCaT cells. Overexpression of EPLIN was found to slow keratinocyte migration rates using two independent assays as well as show convincing association and interaction with two NWASP and E-Cadherin. These important findings suggest novel routes to positively manipulate the wound healing process and has significance in further translational research.