title: Differential expression of proopiomelanocortin (POMC) transcriptional variants in human skin cells.
abstract: The aims of this study were to examine content and expression level of proopiomelanocortin (POMC) mRNA variants in human dermal fibroblasts (HDF) as compared to primary keratinocytes and HaCaT cells of keratinocyte origin. Primary fibroblasts and keratinocytes were obtained from normal human foreskin. Full-length and total (i.e. the full-length, truncated and/or alternatively spliced) POMC mRNA in skin cells were determined by qRT-PCR using specific probes. The full-length POMC mRNA in HDF is neither constitutively expressed, nor could be induced by corticotropin releasing hormone (CRH) or cytokines interferon γ (IFN-γ) and transforming growth factor-β1 (TGF-β1). However, the truncated/alternatively spliced POMC mRNA variants are constitutively expressed in HDF and could be moderately increased with CRH and the cytokines. Primary keratinocytes, in addition to truncated/alternatively spliced POMC mRNA variants, also constitutively express full-length POMC mRNA, both being downregulated during in vitro culturing. Unlike primary keratinocytes, HaCaT cells, express only truncated/alternatively spliced POMC mRNA variants. The level of POMC mRNA expression in HaCaT cells was associated with differentiation stage, being higher in more differentiated cells. Thus, in this study we have shown for the first time that HDF do not express the full-length POMC mRNA, either constitutively or upon activation, opposing to primary keratinocytes which constitutively express the full-length POMC mRNA as a minor variant. Although expressing only truncated/alternatively spliced POMC mRNA variant, HDF express POMC peptide, showing that those transcriptional variants are translatable. Truncated/alternatively spliced POMC mRNA variants, expressed both in HDF and keratinocytes are subjected to regulation, implicating their functionality. Furthermore, the IFN-γ-induced up-regulation at transcriptional level was associated with increased level of POMC peptide detected in HDF lysates. Thus, data of this study have shown that HDF express only truncated/alternatively spliced POMC mRNA variants, which are probably biologically relevant as they could be translated to POMC peptide, both constitutively and upon activation.
authors.affiliation: Ruđer Bošković Institute, Division of Molecular Medicine, Laboratory for Experimental Haematology, Immunology and Oncology, Zagreb, Croatia.
authors.forename: E
authors.initials: E
authors.lastname: Zapletal
authors.forename: O
authors.initials: O
authors.lastname: Kraus
authors.forename: B
authors.initials: B
authors.lastname: Cupić
authors.forename: J
authors.initials: J
authors.lastname: Gabrilovac
mesh_terms: D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D016922:Cellular Senescence / Q000502:physiology; D002648:Child; D002675:Child, Preschool; D003346:Corticotropin-Releasing Hormone / Q000096:biosynthesis; D004797:Enzyme-Linked Immunosorbent Assay; D005347:Fibroblasts; D014644:Genetic Variation; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis; D015603:Keratinocytes / Q000378:metabolism; D008297:Male; D010902:Pituitary Gland / Q000378:metabolism; D011333:Pro-Opiomelanocortin / Q000096:biosynthesis* / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D012867:Skin / Q000166:cytology / Q000378:metabolism*; D014158:Transcription, Genetic; D053773:Transforming Growth Factor beta1 / Q000096:biosynthesis
chemical_list: D012333:RNA, Messenger; D053773:Transforming Growth Factor beta1; D011333:Pro-Opiomelanocortin; D007371:Interferon-gamma; D003346:Corticotropin-Releasing Hormone