This study aims at elucidating the role of cytokines in the mechanism of proliferation of cholesteatoma epithelium by investigating the mode of expression of epidermal growth factors, such as TGF-alpha. The subjects of this study were patients who had undergone operation for middle ear cholesteatoma. Skins of the bone region of the external ear canal (normal skin) of the same patients were used as the negative control. The mode of expression of TGF-alpha was studied by immunohistochemistry and in situ hybridization. In the immunohistochemical study, there were no conspicuous differences observed between cholesteatoma tissues and normal skin. After in situ hybridization, expression of TGF-alpha mRNA was mainly observed in the epidermal basal cell layer in the normal skin, while in the cholesteatoma epidermis with severe inflammatory cell infiltration, expression of TGF-alpha mRNA was observed up to layers superior to the basal cell layer. The expression of TGF-alpha mRNA is greatly affected by subepithelial connective tissue, strongly suggesting involvement of paracrine regulation in proliferation of cholesteatoma epithelium.