Candidate Housekeeping Genes Require Evaluation before their Selection for Studies of Human Epidermal Keratinocytes

Minner, Frédéric; Poumay, Yves

doi:10.1038/jid.2008.247

Cited by 52 publications

(50 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Dnase I-treated (Invitrogen; Life Technologies) and reverse transcription PCR products were analyzed using the MyiQ Single-Color Real-Time Detection System for quantification with SYBR Green and melting curve analysis (Bio-Rad Laboratories). Expression of target genes was normalized to expression of the housekeeping gene human ribosomal phosphoprotein P0 (RPLP0) in the same sample (60). Relative mRNA expression levels were calculated using the ΔΔCt method (61).…”

Section: Figurementioning

confidence: 99%

Coal tar induces AHR-dependent skin barrier repair in atopic dermatitis

Bergboer²,

et al. 2013

View full text Add to dashboard Cite

Topical application of coal tar is one of the oldest therapies for atopic dermatitis (AD), a T helper 2 (Th2) lymphocyte-mediated skin disease associated with loss-of-function mutations in the skin barrier gene, filaggrin (FLG). Despite its longstanding clinical use and efficacy, the molecular mechanism of coal tar therapy is unknown. Using organotypic skin models with primary keratinocytes from AD patients and controls, we found that coal tar activated the aryl hydrocarbon receptor (AHR), resulting in induction of epidermal differentiation. AHR knockdown by siRNA completely abrogated this effect. Coal tar restored filaggrin expression in FLG-haploinsufficient keratinocytes to wild-type levels, and counteracted Th2 cytokine-mediated downregulation of skin barrier proteins. In AD patients, coal tar completely restored expression of major skin barrier proteins, including filaggrin. Using organotypic skin models stimulated with Th2 cytokines IL-4 and IL-13, we found coal tar to diminish spongiosis, apoptosis, and CCL26 expression, all AD hallmarks. Coal tar interfered with Th2 cytokine signaling via dephosphorylation of STAT6, most likely due to AHR-regulated activation of the NRF2 antioxidative stress pathway. The therapeutic effect of AHR activation herein described opens a new avenue to reconsider AHR as a pharmacological target and could lead to the development of mechanism-based drugs for AD.

show abstract

Section: Figurementioning

confidence: 99%

Coal tar induces AHR-dependent skin barrier repair in atopic dermatitis

Bergboer²,

et al. 2013

View full text Add to dashboard Cite

show abstract

“…Primers for CCN transcripts were synthesized according to published sequences (Quan et al 2009). Results are presented as fold change in treated vs. untreated skin sample, normalized to transcript levels of 36B4 (RPLP0, ribosomal protein, large, P0), housekeeping gene (Minner and Poumay 2009). …”

Section: Immunohistochemistry and Imagingmentioning

confidence: 99%

Spatial-temporal modulation of CCN proteins during wound healing in human skin in vivo

Rittié

Perbal

Castellot

et al. 2011

J. Cell Commun. Signal.

View full text Add to dashboard Cite

CCN proteins are important modulators of development and function of adult organs. In this study, we examined the localization and expression of the six CCN family members in normal adult human skin and during wound healing in vivo. Transcript and protein expression were studied by laser-capture microdissection-coupled real-time PCR and immunohistochemistry, respectively. Our results demonstrate that CCN1, CCN4, and CCN6 are expressed at relatively low levels in normal human skin. CCN2, CCN3, and CCN5 are the most highly expressed transcripts in the epidermis. CCN3 and CCN5 proteins are prominent in epidermal keratinocytes, whereas CCN2 is primarily expressed in melanocytes. Differential expression within epidermal layers suggests that CCN3 and CCN5 are linked with keratinocyte differentiation. CCN2, CCN3 and CCN5, are the three most highly expressed transcripts in the dermis. Their respective proteins are produced to various extents by dermal fibroblasts, blood vessels, eccrine sweat glands and hair follicles. We find that most CCN family members are temporally and specifically regulated during different phases (inflammation, proliferation, and remodeling) of partial thickness wound repair. By highlighting spatialtemporal regulations of CCN family member expression in relation to cell proliferation and differentiation, our results suggest a diverse range of functions for CCN proteins in both epidermal and dermal cells, and provides a solid reference for interpretation of future studies aimed at understanding the role of CCN proteins in human skin physiology and diseases.

show abstract

“…In brief, cDNA samples were diluted 1:10 and quantified by amplification against serial dilutions of appropriate control cDNA with the following cycling conditions: initial denaturation 95°C for 10 minutes and 40 cycles of 95°C for 15 seconds, 58°C for 15 seconds, 60°C for 60 seconds. Specific primer sets were utilised for a, b and g p63, p53 (Wang and Seed, 2003), p21Cip1 and RPLPO (large ribosomal protein) (Minner and Poumay, 2009) (for primer sequences, see supplementary material Table S2). Expression levels were assessed in triplicate, normalised to RPLPO levels and graphs represent the combined results of at least three independent biological replicates.…”

Section: Real-time Reverse-transcriptase Pcr Analysismentioning

confidence: 99%

p63 maintains keratinocyte proliferative capacity through regulation of Skp2–p130 levels

McDade

Patel

McCance

2011

Journal of Cell Science

View full text Add to dashboard Cite

p63 is a master regulator of proliferation and differentiation in stratifying epithelia, and its expression is frequently altered in carcinogenesis. However, its role in maintaining proliferative capacity remains unclear. Here, we demonstrate that hypoproliferation and loss of differentiation in organotypic raft cultures of primary neonatal human foreskin keratinocytes (HFKs) depleted of the α and β isoforms of p63 result from p53–p21-mediated accumulation of retinoblastoma (Rb) family member p130. Hypoproliferation in p63-depleted HFKs can be rescued by depletion of p53, p21CIP1 or p130. Furthermore, we identified the gene encoding S-phase kinase-associated protein 2 (Skp2), the recognition component of the SCFSkp2 E3 ubiquitin ligase, as a novel target of p63, potentially influencing p130 levels. Expression of Skp2 is maintained by p63 binding to a site in intron 2 and mRNA levels are downregulated in p63-depleted cells. Hypoproliferation in p63-depleted cells can be restored by re-expression of Skp2. Taken together, these results indicate that p63 plays a multifaceted role in maintaining proliferation in the mature regenerating epidermis, in addition to being required for differentiation.

show abstract

Candidate Housekeeping Genes Require Evaluation before their Selection for Studies of Human Epidermal Keratinocytes

Cited by 52 publications

References 14 publications

Coal tar induces AHR-dependent skin barrier repair in atopic dermatitis

Coal tar induces AHR-dependent skin barrier repair in atopic dermatitis

Spatial-temporal modulation of CCN proteins during wound healing in human skin in vivo

p63 maintains keratinocyte proliferative capacity through regulation of Skp2–p130 levels

Contact Info

Product

Resources

About