Chemical Carcinogenesis Smad3 Knockout Mice Exhibit a Resistance to Skin

It has been shown that Smad3 exerts both tumor-suppressive and -promoting roles. To evaluate the role of Smad3 in skin carcinogenesis in vivo, we applied a chemical skin carcinogenesis protocol to Smad3 knockout mice (Smad3 / and Smad3 / ) and wild-type littermates (Smad3 / ). Smad3 / mice exhibited reduced papilloma formation in comparison with Smad3 / mice and did not develop any squamous cell carcinomas. Further analysis revealed that Smad3 knockout mice were resistant to 12-O-tetradecanoylphorbol-13-acetate (TPA)–induced epidermal hyperproliferation. Concurrently, increased apoptosis was observed in TPAtreated Smad3 / skin and papillomas when compared with those of wild-type mice. Expression levels of activator protein-1 family members (c-jun, junB, junD, and c-fos) and transforming growth factor (TGF)were significantly lower in TPA-treated Smad3 / skin, cultured keratinocytes, and papillomas, as compared with Smad3 / controls. Smad3 / papillomas also exhibited reduced leukocyte infiltration, particularly a reduction of tumor-associated macrophage infiltration, in comparison with Smad3 / papillomas. All of these molecular and cellular alterations also occurred to a lesser extent in Smad3 / mice as compared with Smad3 / mice, suggesting a Smad3 gene dosage effect. Given that TGF1 is a well-documented TPA-responsive gene and also has a potent chemotactic effect on macrophages, our study suggests that Smad3 may be required for TPA-mediated tumor promotion through inducing TGF1– responsive genes, which are required for tumor promotion, and through mediating TGF1–induced macrophage infiltration.