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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Archives of dermatological research 269 (1980), S. 111-126 
    ISSN: 1432-069X
    Keywords: Dermal fibroblasts ; Glucocorticoids ; Receptors ; Growth ; Dermale Fibroblasten ; Glukokorticoide ; Receptoren ; Wachstum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung Es wurden Primärkulturen dermaler Fibroblasten aus neugeborenen Mäusen verwandt, um einige der entzündungshemmenden Wirkungen der Glukocorticoide in vitro unter dem Einfluß des genetischen Hintergrunds zweier verschiedener Mäusestämme (A/J und C57B1/6J) zu untersuchen. Fibroblasten wurden 2-7 Tage lang mit und ohne verschiedene Glukocorticoide kultiviert. Unter Steroiden-4-7 Tage lang — wurde die DNA-Synthese um 50–85% und die Eiweißsynthese um 50–60% gehemmt. Corticosteron verursachte in diesen Zellen eine dosisabhängige Hemmung der DNA-Synthese, wobei eine 50%-Reduzierung bei 10 nM eintrat. Spezifisches Eiweiß mit hoher Affinität und niedriger Kapazität für [3H]-Dexamethasonbzw. [3H]-Triamzinolon-Acetonid wurde im Cytoplasma von Dermalfibroblasten offenbar mit einem Kd von 9 nM und ca. 5200–6400 Bindestellen/Zelle aufgefunden. Die Sedimentationsanalyse des [3H]-Triamzinolon-Acetonid-Receptorenkomplexes wies bei niedrigen Salzglyceringefällen eine Bindung in der 7-8S-Region auf. Diese Untersuchungen weisen darauf hin, daß die Wachstumshemmung von Primärkulturen dermaler Fibroblasten neugeborener Mäuse durch Glukocorticoide wahrscheinlich auf receptoren vermitteltem Wege stattfindet und daß dieses Primärkulturensystem zur Abgrenzung anderer entzündungshemmender Wirkungen, von Glukocorticoiden in vitro nützlich sein dürfte.
    Notes: Summary Primary cultures of dermal fibroblasts from neonatal mice were used to investigate some of the anti-inflammatory effects of glucocorticoids in vitro as influenced by the genetic background of two different strains of mice (A/J and C57 B1/6J). Fibroblasts were cultured in the absence or presence of various glucocorticoids for 2–7 days. After 4–7 days in the presence of steroid, DNA synthesis was reduced by 50–85% while protein synthesis was inhibited by 50–60%. Corticosterone produced a dose-dependent inhibition of DNA synthesis in these cells with a 50% reduction occurring at 10 nM. Specific, high affinity, low capacity binding proteins for [3H]dexamethasone or [3H]triamcinolone acetonide were identified in the cytoplasm of neonatal dermal fibroblasts which had an apparent Kd of 9 nM and ∼5,200–6,400 binding sites/cell. Sedimentation analysis of the [3H]triamcinolone acetonide-receptor complexes on low salt glycerol gradients exhibited binding in the 7 to 8 S region of the gradients. These studies demonstrate that inhibition of growth of primary cultures of mouse neonatal dermal fibroblasts by glucocorticoids is probably mediated by a receptor-mediated pathway, and that this primary culture system might be useful in delineating other anti-inflammatory effects of glucocorticoids in vitro.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 121 (1984), S. 22-30 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Two flat cellular revertant cell lines, F-2 and C-11, which were originally selected from the DT line of Kirsten murine sarcoma virus (Ki-MuSV)-transformed NIH/3T3 cells, were examined for the production of transforming growth factors (TGFs). The revertant cells fail to grow in semisolid medium as colonies and exhibit a markedly reduced level of tumorigenicity in nude mice, although they are known to express high levels of p21ras, the product of the Kirsten sarcoma virus oncogene, ras, and they contain a rescuable transforming virus. TGF activity associated with the transformed, revertant, and non-transformed cell lines was measured by the ability of concentrated conditioned medium (CM) from these cells to induce normal rat kidney (NRK) and NIH/3T3 cells to form colonies in semisolid agar suspension cultures and to inhibit the binding of 125I epidermal growth factor (EGF) to specific cell surface receptors. CM from the transformed DT cells and from both the F-2 and C-11 revertants contains TGF activity, in contrast to CM obtained from normal NIH/3T3 cells. Furthermore, unlike NIH/3T3 cells, neither the DT nor the revertant cells were able to bind 125I EGF. All four cell lines were able to proliferate in serum-free medium supplemented with transferrin, insulin, EGF, and Pedersen fetuin. However, in basal medium lacking these growth factors, only DT cells and, to a lesser extent, the revertant cells were able to grow. These results suggest that the F-2 and C-11 revertants fail to exhibit all of the properties associated with transformation because the series of events leading to the transformed phenotype is blocked at a point(s) distal both to the expression of the p21 ras gene product and also to the production of TGFs and that the production of TGFs may be necessary but not sufficient for maintaining the transformed state.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Epidermal growth factor (EGP) inhibited the growth of A431 human epidermoid carcinoma cells. The tumor promoting, phorbol ester, 12-O-tetradeca-noylphorbol-13-acetate (TPA) also retarded A431 cell growth. Addition of both TPA and ECF inhibited cell growth in an additive or synergistic manner depending upon the initial plating density of the cultures. EGF increased the production of diacylglycerol (60-70%) and stimulated the synthesis of phosphatidylinositol (PI) from 3H-inositol (three- to fourfold increase). Both of these responses were attenuated in the presence of TPA. TPA alone stimulated the production of diacylglycerol (DG) but had little effect on PI synthesis. The biological effect of TPA appeared to be mediated by the presence of a high-affinity receptor for phorbol esters on A431 cells. Moreover, the binding of 125I-EGF to A431 cells was unaffected by TPA, suggesting that the antagonistic effects of TPA were occurring distal to the EGF receptor. These findings also indicated that although TPA and EGF both inhibited A431 cell growth, this effect could be dissociated from changes in PI synthesis but may be dependent upon transient changes in DG production.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
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