Immunohistochemical Analysis of Tumor Tissue

The tumor tissue was taken, fixed with 4% paraformaldehyde solution, dehydrated, and sliced into 4 μm. Sections were routinely dewaxed and hydrated and added with H₂O₂, and microwaves were used to retrieve antigens. The sample was blocked with 10% normal goat serum, and E-cadherin (1:200, American Abgent Company) and N-cadherin (1:200, American Affinity Company) antibodies were added. After the addition of a secondary antibody (Proteintech Company, USA), the sections were developed with diaminobenzidine, followed by staining with hematoxylin and sealing with neutral resin. The stained sections showed brown–yellow granules under the microscope. Ten fields were randomly selected for photography, and analysis of the images was done with Image J software (Media Cybernetics) [36 (link)].