A limited role for retinoic acid and retinoic acid receptors RAR alpha and RAR beta in regulating keratin 19 expression and keratinization in oral and epidermal keratinocytes

Abstract

Different types of stratified squamous epithelia-for example, the "orthokeratinized" epidermis, the "parakeratinized" gingiva, and the "nonkeratinized" oral lining mucosal epithelia-are formed by intrinsically distinct keratinocyte subtypes. These subtypes exhibit characteristic patterns of keratin protein expression in vivo and in culture. Keratin 19 is an informative subtype-specific marker because the basal cells of only nonkeratinizing epithelia express K19 in vivo and in culture. Epidermal keratinocytes normally do not express K19, but can be induced to do so in culture by retinoic acid (RA). Keratinocyte subtypes express the retinoic acid receptor (RAR) beta at levels roughly correlated with their level of K19 expression in culture and their potential for forming a nonkeratinized epithelium in vivo. We tested the hypothesis that the level of RAR beta expressed by a keratinocyte determines its K19 expression and its form of suprabasal differentiation. Normal human epidermal and gingival keratinocytes stably overexpressing either RAR beta or RAR alpha were generated by defective retroviral transduction. Overexpression of either receptor enhanced the RA inducibility of K19 in conventional culture, in that the proportion of the transductants becoming K19+ in response to RA was markedly increased compared with controls. The pattern of differentiation of the epithelium formed in organotypic culture, assessed by basal K19 and suprabasal K1, K4, and filaggrin expression, however, was unaltered by RAR overexpression. Thus, the susceptibility of keratinocytes to regulation of K19 expression by retinoids is conditional, and levels of neither RAR beta nor RAR alpha are limiting to the intrinsic mechanism that specifies alternate differentiation pathways for stratified squamous epithelia.