Effect of Cellular Senescence and Retinoic Acid on the Expression of Cellular Retinoic Acid Binding Proteins in Skin Fibroblasts

Abstract

Retinoic acid (RA) has significant effects on a variety of cellular processes, including growth and differentiation. Retinoic acid has also been implicated as a major morphogen during embryogenesis. In the skin, both the epidermis and the dermis are extremely responsive to the effects of retinoids; however, the molecular mechanisms through which retinoids act in this tissue remain poorly understood. Two classes of proteins play roles in mediating the biological effects of retinoic acid. The nuclear receptors for retinoic acid are of two types, the initially described RARs, and the RXRs. Each of these nuclear receptor families has multiple isoforms and can function as a ligand-inducible transcription enhancing factor. A second class of receptor proteins exists for retinoic acid that are found in the cytoplasm, the cellular retinoic acid binding proteins (CRABPs). These proteins play a role in the binding, transport, and metabolism of retinoic acid. In previous studies, we have demonstrated that retinoic acid induces the expression of RARβ and RARγ in human dermal fibroblasts. Moreover, we have also shown the selective transcriptional up-regulation of the RARβ2 isoform in senescent dermal fibroblasts and senescent human mammary epithelial cells. In order to further define molecules important in regulating the response of senescent dermal fibroblasts to retinoids, we demonstrate here that retinoic acid induces CRABP-II messenger RNA in human dermal fibroblasts in a dose-dependent manner. Moreover, we show that the induction by RA can be inhibited by actinomycin D, suggesting that the up-regulation may be mediated by a transcriptional mechanism. We further demonstrate that cycloheximide also has an effect on the up-regulation, suggesting a role for protein synthesis in the regulation of CRABP-II gene expression. We show that CRABP-II is a very stable messenger RNA species, in contrast to the mRNAs for RARα, RARβ, and RARγ. Of interest, we demonstrate no significant difference in the expression of CRABP-II between presenescent and senescent fibroblasts. Taken together, these data suggest that retinoic acid plays a central role in the regulation of CRABP-II gene expression in the dermal fibroblast and that this molecule is the major mediator of the cytoplasmic effects of retinoids in dermal fibroblasts. However, in contrast to RARβ2, there is no apparent change in the regulation of CRABP-II in senescent dermal fibroblasts.