Phosphorylated ezrin is associated with EBV latent membrane protein 1 in nasopharyngeal carcinoma and induces cell migration

Tumor metastasis is a complex phenomenon that is the culmination of effects of numerous cellular factors. We have shown that the EBV oncoprotein, latent membrane protein 1 (LMP1), is capable of inducing a wide range of such factors in cell culture, expression of which is also elevated in the LMP1-expressing tumor, nasopharyngeal carcinoma (NPC), a highly invasive neoplasm. Recently, the membrane-crosslinker protein ezrin has been implicated in tumor cell metastasis and malignant progression. In this study, we evaluated the possible role of LMP1 and ezrin in the pathophysiology of NPC. We show that C-terminal phosphorylation of ezrin is increased by expression of LMP1 in nasopharyngeal (NP) cells through a Protein Kinase C (PKC) pathway. LMP1 enhances organization of a ternary complex of CD44, ezrin and F-actin which is a prerequisite for ezrin phosphorylation. In NPC tissues, expression of phosphoezrin and LMP1 is directly correlated. Silencing of endogenously expressed ezrin suppresses LMP1-induced cell motility and invasiveness. Moreover inhibition of ezrin phosphorylation by PKC inhibitor suppresses migration and invasion of NP cells. These data demonstrate that phosphorylation of ezrin and its recruitment to the cell membrane linked to F-actin and CD44 is a process required for LMP1-stimulated cell motility and invasion of NP cells.