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The retinoid signalling molecule, tripartite motif protein 16 (TRIM16), is a regulator of keratinocyte differentiation and a tumour suppressor in retinoid\u2010sensitive neuroblastoma. We sought to determine the role of TRIM16 in skin squamous cell carcinoma (SCC) pathogenesis. We have shown that TRIM16 expression was markedly reduced during the histological progression from normal skin to actinic keratosis and SCC. SCC cell lines exhibited lower cytoplasmic and nuclear TRIM16 expression compared with primary human keratinocyte (PHK) cells due to reduced TRIM16 protein stability. Overexpressed TRIM16 translocated to the nucleus, inducing growth arrest and cell differentiation. In SCC cells, TRIM16 bound to and down regulated nuclear E2F1, this is required for cell replication. Retinoid treatment increased nuclear TRIM16 expression in retinoid\u2010sensitive PHK cells, but not in retinoid\u2010resistant SCC cells. Overexpression of TRIM16 reduced SCC cell migration, which required the C\u2010terminal RET finger protein (RFP)\u2010like domain of TRIM16. The mesenchymal intermediate filament protein, vimentin, was directly bound and down\u2010regulated by TRIM16 and was required for TRIM16\u2010reduced cell migration. Taken together, our data suggest that loss of TRIM16 expression plays an important role in the development of cutaneous SCC and is a determinant of retinoid sensitivity. Copyright \u00a9 2011 Pathological Society of Great Britain and Ireland. 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