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  • CELL  (31)
  • KERATINOCYTES  (16)
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  • 1
    Keywords: EXPRESSION ; IN-VITRO ; DISTINCT ; DIFFERENTIATION ; MESSENGER-RNA ; murine ; KERATINOCYTES ; EPIDERMAL DIFFERENTIATION ; ARACHIDONIC-ACID ; calcium gradient ; ENHANCING FACTOR ; epidermal barrier ; GROUP-II ; GROUP-V ; GROUP-X ; hyperproliferation ; INVITRO CULTIVATION ; LOW-MOLECULAR-WEIGHT ; neonatal mouse ; NEONATAL MOUSE KERATINOCYTES ; PERMEABILITY BARRIER HOMEOSTASIS ; epidermis
    Abstract: The action of secreted phospholipases A(2) in skin is thought to be essential for epidermal barrier homeostasis. The incomplete knowledge of presence and functions of the novel secreted phospholipase A(2) subtypes in skin prompted us to explore their expression in epidermis and primary keratinocytes from murine neonatal skin. We detected secreted phospholipases A(2) -IB, -IIA, -IIC, -IID, -IIE, -IIF, -V, -X, and -XII. To study secreted phospholipase A(2) expression during epidermal differentiation, primary keratinocytes from the basal, suprabasal, and upper differentiated layers of neonatal mouse epidermis were obtained by density gradient centrifugation. mRNA for secreted phospholipases A(2) -IB, -IIE, -IIF, -V, and -XII-1 are mainly expressed in the upper differentiated layers, whereas the most prominent enzymes in the basal and suprabasal layers are secreted phospholipases A(2) -IIA, -IID, and -X. The mRNA for secreted phospholipase A(2) -IIC was found in all fractions. Immunohistochemical analysis in mouse skin sections reflected the mRNA distribution patterns in the different epidermal cell fractions. After in vitro induction of keratinocyte differentiation by increasing the calcium concentration of the medium, secreted phospholipases A(2) -IB, -IIE, -IIF, -V, and -XII-1 were upregulated, whereas secreted phospholipases A(2) -IIA, -IIC, -IID, and -X were mainly expressed in proliferating keratinocytes. The specific secreted phospholipase A(2) expression profile in the skin suggests a distinct function for each enzyme in the epidermis
    Type of Publication: Journal article published
    PubMed ID: 12839576
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  • 2
    Keywords: CELLS ; EXPRESSION ; IN-VITRO ; INHIBITOR ; proliferation ; CELL ; CELL-PROLIFERATION ; Germany ; human ; GENERATION ; SYSTEM ; DISTINCT ; PROTEIN ; PROTEINS ; cell line ; LINES ; ACTIVATION ; RESPONSES ; REDUCTION ; KERATINOCYTES ; SKIN ; CELL-LINES ; ISOFORM ; SUBUNIT ; Western-blot ; MEMBRANE ; CELL-LINE ; LINE ; CYTOCHROME-C ; EPITHELIAL-CELLS ; PROTEIN LEVELS ; western blot ; HaCaT ; MUCOSA ; HOST-DEFENSE ; DEFENSE ; human skin and oral epithelial cells,oxidoreductase,p67phox,spin trapping,superoxide radical ; NAD(P)H OXIDASE ; OXYGEN RADICALS ; P47(PHOX) ; SUPEROXIDE-PRODUCTION
    Abstract: In non-phagocytic cells, superoxide has been implicated in physiological and pathological cellular functions in the skin and mucosa, such as, host defense, mitogenic responses, and malignant conversion. Here, we identify a constitutively expressed heme-flavoprotein NADPH oxidase (Nox) system as a source of superoxide in human skin (HaCaT) and gingival mucosal (GM16) keratinocyte cell lines. Western blot analysis showed that both cell lines expressed the phagocyte oxidase (phox) cytosolic proteins Rac1, p40phox, and p67phox. With respect to the catalytic flavoheme protein subunit, HaCaT membranes, which expressed p22phox, showed an absorbance peak at 558 nm indicative of a b-type cytochrome. At mRNA levels, both GM16 and HaCaT cells expressed gp91phox homologs Nox1, Nox2, and Nox4, however, HaCaT cells expressed very low levels of Nox1 mRNA. At protein levels, Nox1 was readily detected in HaCaT but was nearly undetectable in GM16 cells. Consistently, Nox activity of HaCaT membranes was demonstrated by electron paramagnetic resonance spin-trapping and cytochrome c reduction, and the activity was sensitive to the flavoprotein inhibitor diphenylene iodonium. V-max values were 20-fold lower than those reported for phagocytic oxidase. In conclusion, keratinocytes expressed a Nox distinct from the phox isoform of phagocytes providing molecular evidence for a source of superoxide that may regulate cell proliferation and host defense in skin and oral mucosa
    Type of Publication: Journal article published
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  • 3
    Keywords: CELLS ; EXPRESSION ; tumor ; CELL ; Germany ; human ; PATHWAY ; GENE ; GENE-EXPRESSION ; GENES ; DIFFERENTIATION ; TUMORS ; COMPLEX ; COMPLEXES ; INDUCTION ; CONTRAST ; SKIN ; LOCALIZATION ; BENIGN ; keratin ; skin tumors ; epidermis ; FOLLICLE ; HAIR-FOLLICLES ; HUMAN TYPE-I ; MATRIX ; BETA-CATENIN EXPRESSION ; CORTEX ; HAIR FOLLICLE ; hair follicles,human,transcription factors,tumors ; HOXC13 ; INVOLUCRIN
    Abstract: Human hair follicles exhibit a complex pattern of sequential hair keratin expression in the hair matrix, cuticle, and cortex. In pilomatricomas, that is, benign skin tumors thought to arise from germinative matrix cells of the hair follicle and retaining morphological signs of cortical differentiation, this differential hair keratin pattern has been shown to be faithfully preserved in the lower and upper transitional cell compartments of the tumors. Here we show that also the co-expression of hair keratin hHa5 with its regulatory nuclear homeoprotein HOXC13 in matrix cells of the hair follicle is maintained in lower transitional cells of pilomatricomas. In contrast, the nuclear co-expression of LEF1 and beta-catenin, which in the hair follicle has been postulated to initiate cortex cell differentiation through the induction of hair keratin hHa1 expression (Merill et al, Genes Dev 15:1688-1705, 2001), is not preserved in upper transitional cells of pilomatricomas. Although these cells correctly express hHa1, they are completely devoid of LEF1 and nuclear LEF1/beta-catenin co-expression is shifted to a subpopulation of hair keratin-free basaloid cells of the tumors. These data imply that unlike the normal hair follicle, cortical differentiation in pilomatricomas is not under the control of the canonical Wnt signaling pathway
    Type of Publication: Journal article published
    PubMed ID: 15140206
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  • 4
    Keywords: RECEPTOR ; APOPTOSIS ; EXPRESSION ; IN-VITRO ; proliferation ; CELL ; Germany ; INHIBITION ; VITRO ; SYSTEM ; DEATH ; PROTEIN ; PROTEINS ; DRUG ; DIFFERENTIATION ; EPITHELIA ; ACTIVATION ; MARKER ; CONTRAST ; SKIN ; treatment ; ALPHA ; culture ; MATURATION ; CELL-DEATH ; ADHESION ; CELL-ADHESION ; RECEPTORS ; PROGRAMMED CELL-DEATH ; BARRIER FUNCTION ; TERMINAL DIFFERENTIATION ; AUTOIMMUNITY ; FUNCTIONAL-CHARACTERIZATION ; cell adhesion ; DRUGS ; ORGANOTYPIC COCULTURE ; cholinergic ; DARIERS-DISEASE ; KERATINOCYTE ADHESION ; NICOTINIC ACETYLCHOLINE-RECEPTOR ; PEMPHIGUS-VULGARIS
    Abstract: The aim of this study was to analyze the influence of cholinergic and anticholinergic drugs on epidermal physiology using organotypic cocultures (OTCs). Blocking of all acetylcholine receptors (AChRs) by combined treatment with mecamylamine and atropine or treatment with strychnine (blocking alpha 9nAChR) for 7-14 days resulted in a complete inhibition of epidermal differentiation and proliferation. Blockage of nicotinic (n) AChR with mecamylamine led to a less pronounced delay in epidermal differentiation and proliferation than blockage of muscarinic ( m) AChR with atropine, evidenced by reduced epithelial thickness and expression of terminal differentiation markers like cytokeratin 2e or filaggrin. In OTCs treated with atropine, mecamylamine, or strychnine, we could demonstrate intracellular lipid accumulation in the lower epidermal layers, indicating a severely disturbed epidermal barrier. In addition, we observed prominent acantholysis in the basal and lower suprabasal layers in mecamylamine-, atropine-, and strychnine-treated cultures, accompanied by a decreased expression of cell adhesion proteins. This globally reduced cell adhesion led to cell death via intrinsic activation of apoptosis. In contrast, stimulation of nAChR and mAChR with cholinergic drugs resulted in a significantly thickened epithelium, accompanied by an improved epithelial maturation. In summary, we show that epidermal AChR are crucially involved in the regulation of epidermal homeostasis
    Type of Publication: Journal article published
    PubMed ID: 16810300
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  • 5
    Keywords: CANCER CELLS ; CELLS ; EXPRESSION ; tumor ; TUMOR-CELLS ; CELL ; CLINICAL-TRIAL ; COMBINATION ; evaluation ; Germany ; human ; IN-VIVO ; MODEL ; THERAPY ; NEW-YORK ; EFFICIENCY ; TRANSDUCTION ; primary ; prognosis ; DOMAIN ; culture ; GLYCOPROTEIN ; virus ; TRIAL ; TRIALS ; VECTORS ; VECTOR ; MEMBRANE ; CLINICAL-TRIALS ; chemotherapy ; EFFICIENT ; MELANOMA ; MALIGNANT-MELANOMA ; malignant melanoma ; CUTANEOUS MELANOMA ; ADENOVIRUS ; DACARBAZINE ; DOMAINS ; THERAPIES ; MELANOMA-CELLS ; VIROTHERAPY ; USA ; EFFICIENT TRANSDUCTION ; SHORT-TERM ; xenograft ; clinical trial ; ONCOLYTIC ADENOVIRUSES ; B ADENOVIRUSES ; CELLULAR RECEPTOR ; FUSOGENIC MEMBRANE-GLYCOPROTEINS ; REPLICATING ADENOVIRUS ; SUICIDE GENE-THERAPY ; ADENOVIRUS VECTORS ; IMMUNE-MEDIATED CONTROL ; oncolytic adenovirus
    Abstract: Advanced melanoma is associated with poor prognosis warranting the development of new therapeutics, such as oncolytic adenoviruses for immunovirotherapy. Since this approach critically depends on efficient transduction of targeted tumor cells, we screened a panel of 22 different adenovirus types for their internalization efficiency in melanoma cells. We demonstrated that the virions of Ad35, Ad38, and Ad3 have significantly higher internalization efficiency in melanoma cells than Ad5, so far the only adenovirus type used in clinical trials for melanoma. Therefore, we developed a conditionally replication-competent Ad5-based vector with the Ad35 fiber shaft and knob domains (Ad5/35) and compared its therapeutic efficacy with the homologous vector carrying the native Ad5 fiber. To further enhance virotherapy, we combined the oncolytic adenovirus vectors with intratumoral expression of measles virus fusogenic membrane glycoproteins H and F (MV-H/F) and dacarbazine chemotherapy. In a human melanoma xenograft model, established from a short-term culture of primary melanoma cells, we demonstrated that the Ad5/35-based therapy had a significantly greater anti-neoplastic effect than the homologous Ad5-based therapy. Furthermore, the combination of virotherapy, intratumoral expression of MV-H/F, and chemotherapy was clearly superior to single- or double-agent therapy. In conclusion, Ad35-based vectors are promising for the treatment of melanoma
    Type of Publication: Journal article published
    PubMed ID: 17960177
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  • 6
    Keywords: RECEPTOR ; CELLS ; EXPRESSION ; GROWTH ; CELL ; Germany ; human ; MODEL ; GENE ; GENES ; transcription ; LIGAND ; SKIN ; BIOLOGY ; fibroblasts ; TARGET ; IN-SITU ; MONOCLONAL-ANTIBODIES ; EPITHELIAL-CELLS ; INDIVIDUALS ; TARGETS ; RECEPTORS ; DISSECTION ; SERUM ; mRNA ; hair ; USA ; THYROTROPIN RECEPTOR ; HPA axis ; CONNECTIVE-TISSUE ; CORTICOTROPIN-RELEASING HORMONE ; FUNCTIONAL-ROLE ; PIGMENTARY UNIT ; SMOOTH MUSCLE ACTIN ; TSH RECEPTOR
    Abstract: Pituitary thyroid-stimulating hormone (TSH) regulates thyroid hormone synthesis via receptors (TSH-R) expressed on thyroid epithelial cells. As the hair follicle (HF) is uniquely hormone-sensitive and, hypothyroidism with its associated, increased TSH serum levels clinically can lead to hair loss, we asked whether human HFs are a direct target for TSH. Here, we report that normal human scalp skin and microdissected human HFs express TSH-R mRNA. TSH-R- like immunoreactivity is limited to the mesenchymal skin compartments in situ. TSH may alter HF mesenchymal functions, as it upregulates alpha-smooth muscle actin expression in HF fibroblasts. TSH-R stimulation by its natural ligand in organ culture changes the expression of several genes of human scalp HFs (for example keratin K5), upregulates the transcription of classical TSH target genes and enhances cAMP production. Although the functional role of TSH in human HF biology awaits further dissection, these findings document that intracutaneous TSH-Rs are fully functional in situ and that HFs of female individuals are direct targets for nonclassical, extrathyroidal TSH bioregulation. This suggests that organ-cultured scalp HFs provide an instructive and physiologically relevant human model for exploring nonclassical functions of TSH, in and beyond the skin
    Type of Publication: Journal article published
    PubMed ID: 19052559
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  • 7
    Keywords: CELLS ; tumor ; CELL ; human ; COMMON ; DISEASE ; SITES ; PROTEINS ; SAMPLE ; SAMPLES ; TUMORS ; TIME ; PATIENT ; DNA ; SKIN ; papillomavirus ; antibody ; IN-SITU ; LESIONS ; COPY NUMBER ; human papillomavirus ; GENOTYPES ; HPV ; REPLICATION ; glutathione-S-transferase ; PSORIASIS ; EPIDERMODYSPLASIA-VERRUCIFORMIS ; hair ; GENOTYPE ; NONMELANOMA SKIN-CANCER ; USA ; PLUCKED EYEBROW HAIRS ; CLINICAL-ASPECTS ; HAIRS ; HUMAN-PAPILLOMAVIRUS-DNA
    Abstract: Epidermodysplasia verruciformis (EV) is a rare disease, characterized by cutaneous warts and associated with a strong predisposition to beta-genus human papillomavirus (HPV). Earlier studies reported high copy numbers of HPV-DNA in nearly all skin tumors from EV patients, but neither HPV replication status in non-lesional skin nor anti-HPV seroreactivity in these patients have been reported yet. We therefore performed a comprehensive viral load analysis for the more common beta-HPV types on skin samples and plucked eyebrow hairs from four EV patients treated at our dermatology department. The results clearly demonstrate that they carry a multiplicity (up to eighteen types) of beta-HPV genotypes in both skin sites. Worthy of note, a high intrapatient concordance for specific types between hair bulbs and skin biopsies was observed and the same beta-PV profile was maintained over time. Viral load analysis revealed a load range between less than one HPV-DNA copy per 100 cells to more than 400 HPV-DNA copies per cell in both eyebrow hairs and skin proliferative lesions. Evaluation of seroreactivity to beta-HPV types in the four EV patients revealed that antibodies against the 16 beta-HPV were significantly more prevalent and showed higher titers than in the controls
    Type of Publication: Journal article published
    PubMed ID: 18923444
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  • 8
    Keywords: CELLS ; EXPRESSION ; GROWTH ; IN-VITRO ; proliferation ; SURVIVAL ; CELL ; CELL-PROLIFERATION ; Germany ; IN-VIVO ; VIVO ; microarray ; RNA ; ADHESION MOLECULES ; MOLECULES ; TISSUE ; SUPPRESSION ; BREAST-CANCER ; TARGET ; CELL-SURVIVAL ; PROGRESSION ; METASTASIS ; MELANOMA ; ADHESION ; MIGRATION ; EPITHELIAL-CELLS ; L1 ; MALIGNANT-MELANOMA ; TARGETS ; CELL-ADHESION MOLECULE ; OVEREXPRESSION ; DIFFERENTIAL EXPRESSION ; AMYLOID PRECURSOR PROTEIN ; chemoresistance ; CELL-GROWTH ; E-cadherin ; development ; tissue microarray ; ALPHA-SECRETASE
    Abstract: ADAM10 (a disintegrin and metalloproteinase 10) is involved in the ectodomain shedding of various substrates, including adhesion molecules such as L1 cell adhesion molecule (L1-CAM) and CD44, which are known to have important roles in the development of malignant melanoma. In our Study, we characterized the expression of ADAM10 in melanoma cells in vitro and in vivo Immunohistochemical analysis oil tissue microarrays indicated that ADAM-10 expression was significantly elevated in melanoma metastasis compared with primary melanomas. In vitro downregulation of ADAM10 with specific small interfering RNA (siRNA) resulted in a suppression of the anchorage-independent cell growth and reduced the migration of melanoma cells. In addition, overexpression of ADAM-10 induced the migration of melanoma cells. In cell lines from melanoma patients with metastasis, ADAM10 was significantly overexpressed, and ADAM10 expression correlated with increased cell proliferation. Furthermore, we present evidence that ADAM-10 is involved in the release of L1-CAM from melanoma cells. It is important that knockdown of cellular L1-CAM reduced the migration of melanoma cells and abrogated the chemoresistance against cisplatin. In contrast, soluble L1-CAM had no effect on melanoma cell migration or cell survival. Taken together, Our data demonstrate that ADAM10 and L1-CAM have important roles during melanoma progression and both molecules represent attractive targets for therapeutical intervention of melanomas
    Type of Publication: Journal article published
    PubMed ID: 19865098
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  • 9
    Keywords: CELLS ; ACTIVATION ; KERATINOCYTES ; SKIN ; CYCLE ; MIGRATION ; E6 ; E-cadherin ; EPIDERMODYSPLASIA-VERRUCIFORMIS ; LIFE ; DNA LOADS
    Abstract: Human papillomaviruses (HPVs), which are contained in the alpha, beta, gamma, mu, and nu genera, differ in their oncogenic potential and their tropism for cutaneous or mucosal epidermis. Langerhans cells (LC), the only epidermal professional antigen-presenting cells, are readily detected in normal mucosal and cutaneous epithelium. The aim of this study is to determine whether LC loss, which has been reported for HPV16, occurs in other HPV genera and establish its significance in viral pathology. We found that, as for HPV16, LCs were reduced in lesions infected with high-risk mucosal (alpha 7 and alpha 9 species) and low-risk cutaneous (gamma and mu) types. Lesions infected with alpha 10 low-risk genital types had reduced LC but contained epidermal LC patches, coincident with dermis-localized regulatory T cells (T-regs). In contrast to other genera, LCs were common in the epidermis, and T-regs occupied the dermis of the potentially high-risk cutaneous beta-HPV type infected lesions. Therefore, LC loss in the infected lesions occurred irrespective of tropism or oncogenic potential of the HPV type. LC depletion in the HPV-infected epidermis may create an environment that is permissive for viral persistence and in HPV lesions in which LCs are found, the presence of typically immunosuppressive T-regs may compensate for their continued presence
    Type of Publication: Journal article published
    PubMed ID: 19759549
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  • 10
    Keywords: EXPRESSION ; AGENTS ; human ; GENE ; PROTEIN ; DIFFERENTIATION ; TISSUE ; MICE ; MECHANISM ; INDUCTION ; TISSUES ; KERATINOCYTES ; mechanisms ; SKIN ; SEQUENCE ; SEQUENCES ; STAGE ; TRANSGENIC MICE ; IDENTIFICATION ; PROMOTER ; transgenic ; REGION ; FRANCE ; hyperproliferation ; epidermis ; TERMINAL DIFFERENTIATION ; LAYER ; MAMMALIAN-TISSUES ; HASSALLS CORPUSCLES ; FOLLICLE ; HAIR-FOLLICLES ; HUMAN TISSUES ; INNER-ROOT-SHEATH ; AGENT ; PATTERN ; HAIR FOLLICLE ; corneodesmosin ; CORNIFIED EPITHELIA ; GENE PROMOTER ; hyperkeratosis ; KERATINOCYTE DIFFERENTIATION ; promoter regions ; PSORIASIS SUSCEPTIBILITY ; REPORTER GENE ; S GENE ; STRATUM-CORNEUM
    Abstract: Corneodesmosin (CDSN) is a desmosomal protein expressed in the epidermis during the late stages of differentiation and in the inner root sheath of hair follicles. The homophilic adhesive properties of the protein suggest that it reinforces keratinocyte cohesion in the upper layers of the epidermis (stratum granulosum and stratum corneum). In this study, we analyzed the expression of the CDSN gene in 16 human tissues. We confirmed the closely restricted expression pattern of CSDN. Indeed, apart from the skin, the mRNA was significantly detected only in the placenta and the thymus. As a step in elucidating the mechanisms of tissue-specific expression, transgenic mice bearing a 4.2 kb fragment of the human CSDN gene promoter linked to the LacZ gene were generated. The reporter-gene expression was detected in special areas of the inner root sheath of the hair follicles and the hair medulla but not in the epidermis. Induction of epidermis hyperproliferation however either by pharmacological agents or by wounding led to strong expression of the reporter gene in the keratinocytes of the stratum granulosum and the parakeratotic corneocytes of the stratum corneum. The data suggest that the genomic sequences and/or regulating factors responsible for the cell-specific expression of the human CDSN gene in the normal hair follicle as well as in the hyperproliferative epidermis are different from those necessary for expression in the normal epidermis
    Type of Publication: Journal article published
    PubMed ID: 15086560
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