Three-dimension cell culture in three HNSCC cell lines (Hep-2, Tb, and CNE-2) ((a), (b), and (c)). (a) The protein expression of EphA2 was inhibited by siRNA and retrieved after a sequence of overexpression of EphA2 was transfected into siEphA2 cells, which were monitored by Western blotting. (b) Disruption of EphA2 by siRNA leading to the change expression of EMT-related molecules measured by Western blotting. There was a decreasing expression of Twist and Vimentin and an increasing expression of E-cadherin, claudin4, and DSG-3 compared with the control cells (P < 0.05). (c) Hep-2, Tb, and CNE-2 cell lines can construct channel-like structures in Matrigel. All cell lines lost the capability to form channel-like structures in Matrigel after EphA2 was knocked down compared with the control cells. And regaining of EphA2 expression reconstructed channel-like networks formation in three cell lines. The black arrows represent the cross-section of the tubular networks, indicating that they contain hollow lumen-like structures (original magnification: ×400).