2.1.Immunohistochemistry:
All tumor specimens were fixed in 10% buffered formalin and embedded in parafin according to standard procedures. Serial 5µm- thick sections were placed on positively charged slides. Immunohistochemical (IHC) detection of E- cadherin (Mouse mAb clone NCH-38, IR05961-2, DAKO), β- catenin (Mouse mAb clone beta-cat-1, IR70261-2, DAKO), N- cadherin (Mouse mAB clone D-4, sc-8424, Santa Cruz, CA, USA), smooth muscle actin (Mouse mAb, Clone 1A4, DAKO), and Zeb1 (Rabbit pAb, ab87280, ing ABCAM) were performed. All IHC stainings were performed by an autostainer Link AS48 (DAKO, Denmark) which uses the envision flex system. The sections were observed and photographed using an Nicon Eclipse Ni microscope equipped with a Nicon digital sight DS-U3 camera.