| Objective:It has been shown that EphA2overexpression promotes human glioblastomas and prostate cancer increased tumor cell invasion and metastasis, and plays a role in maintaining the characteristics of cancer stem cells. We previuosly has established a nasopharyngeal carcinoma (NPC)5-8F cell line that overexpressed EphA2with S897mutation (S897A-EphA2), and in this study we explore the effects of897phosphorylation of EphA2on tumor cell invasion and metastasis, as well as cancer stem cell characteristics in NPC.Methods:Using our establishing5-8F cell line that overexpressed EphA2with S897mutation(5-8F-EphA2-S897A), and5-8F cell line transfected with empty vector (5-8F-vector), and untransfected5-8F as samples, we performed the following experiments:(1) scratch wound-healing assay and transwell cell invasion assay to detect in vitro cell migratory and invasive ability;(2) phase contrast microscope was used to observe TGF-α-induced cell morphological changes, and Western blot was performed to detect the expression of EMT marker proteins(E-cadherin and Vimentin);(3) sorting of flow cytometry to determine the proportion of side population cells, and tumor sphere culture was performed to observe the capabilities of tumor sphere formation;(4) experimental lung metastasis of nude mice was used to determine the capabilities of tumor cell metastasis.Results:Compared with control cells (5-8F-vector cells and5-8F cells),5-8F-EphA2-S897A cells possessed the following characteristics:(1) the mobility of in vitro cell migration and invasion was significantly decreased;(2) TGF-α induced EMT was inhibited;(3) the ratio of side population cells and the capability of tumor sphere formation significantly reduced;(4) the number and size of lung metastatic tumors were significantly decreased.Conclusions:Serine897phosphorylation of EphA2enhances the ability of tumor cell invasion and metastasis, induces EMT, and maintains the characteristics of cancer stem cell in NPC cells. |
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