Plasminogen activator in cultured Lewis lung carcinoma cells measured by chromogenic substrate assay

Whur, P; Magudia, M; Boston, Julie; Lockwood, Julia; Williams, Donald C.

doi:10.1038/bjc.1980.231

Cited by 26 publications

(11 citation statements)

References 14 publications

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“…purchased from Gibco (BRL, Grand Island, N.Y.), supplemented with 15% fetal bovine serum on gela tin-coated dishes and treated for 3 days with retinoids as described by Strickland and Mahdavi [ 11], Differ entiation was estimated by microscopical observation o f morphological changes and was quantified by as saying PA secretion as described [56]. The results were normalized according to cellular protein con tent.…”

Section: Introductionmentioning

confidence: 99%

Biological Activity of Retinoids Correlates with Affinity for Nuclear Receptors but Not for Cytosolic Binding Protein

Darmon¹,

Rocher²,

Cavey³

et al. 1988

Skin Pharmacol Physiol

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In order to better understand the respective roles of the nuclear retinoic acid receptors (RARs) and the cytosolic retinoic acid binding protein (CRABP) in the mode of action of retinoic acid (RA), several types of RA analogs have been synthesized. Representative compounds have been radiolabeled to a high specific activity and their binding (direct and competition) to RARs and CRABP was determined. Their biological activity on F9 embryonal carcinoma cell differentiation has been determined by a quantitative assay of plasminogen activator (PA). All biologically active analogs studied in this work bound to RARs. A good correlation was found between PA induction and affinity for the RARs, with the exception of RA itself which was a good ligand but a moderate inducer of F9 differentiation. Two biologically active analogs (compounds II and III) did not bind to the CRABP. One biologically inactive analog (compound VIII) bound to CRABP. These results strongly suggest that retinoids must bind to RARs but not necessarily to CRABP in order to induce cell differentiation in F9 cells.

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Section: Introductionmentioning

confidence: 99%

Biological Activity of Retinoids Correlates with Affinity for Nuclear Receptors but Not for Cytosolic Binding Protein

Darmon¹,

Rocher²,

Cavey³

et al. 1988

Skin Pharmacol Physiol

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“…Plasminen activator Plasminogen activator activity was masured by a chromogenic assay which cleaves p-nitroanilie from a conjugated peptide S-2251 (Kabi-Vitrum) in the presnce of 0.15 mg ml-l poly-D-lysine (Whur et al, 1980). The yellow product was read on an ELISA plate reader at 405 nm.…”

mentioning

confidence: 99%

Activity of interferon α, interleukin 6 and insulin in the regulation of differentiation in A549 alveolar carcinoma cells

McCormick

Freshney²,

Speirs³

1995

Br J Cancer

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S_umary The differentiation of A549. a human tumour cell line from type II pneumocytes. can be induced by a crude fibroblast-derived factor (FDF) isolated from the conditioned medium of glucocorticoid-treated lung fibroblasts. In the present report. we have used alkaline phosphatase as a differentiation marker to investigate the activity of a number of growth factors as potential candidates for this paracrine activity. This showed that insulin. interleukin 6 (IL-6), and interferon m (IFN-a) Edelson et al. (1988) showed that maturation of the type II pneumocyte is accompanied by an increase in alkaline phosphatase (AP) activity, and this marker has been used in the present study to demonstrate that conditioned medium from DX-treated fibroblasts (DXCM) also induces AP activity in A549 cells. DX is active on A549 cells alone, but its activity is greatly enhanced by conditioned medium from DX-treated lung fibroblasts. However, while this conditioned medium is active for the induction of PS synthesis after DX had been removed (Speirs et al., 1991), the continued presence of DX is required for induction of AP activity. The precise role of DX is as yet unclear.The

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“…Plasminogen activator (PA) A modification of the chromogenic assay developed by Whur et al (1980) was used to determine PA activities. Cells were washed 3 times with PBSA and incubated with a solution containing HBSS (without phenol red) (Flow Laboratories), glucose (0.1%) and vitamins, 1mM chromogenic substrate S-2302 (Kabivitrum), 1 caseine unit ml-1 plasminogen (Kabivitrum) and 0.15mgmlP1 poly-D-lysine (Sigma).…”

Section: Methodsmentioning

confidence: 99%

Interrelationship between differentiation and malignancy-associated properties in glioma

Frame¹,

Freshney²,

Vaughan³

et al. 1984

Br J Cancer

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Summary The phenotypic expression of cells derived from human anaplastic astrocytomas, rat glioma, normal human adult and foetal brain tissue have been examined for differentiated and malignancy-associated properties. Glial fibrillary acidic protein (GFAP), high affinity glutamate and y-amino butyric acid (GABA) uptake and glutamine synthetase were used as indicators of astroglial differentiation. Plasminogen activator and tumour angiogenesis factor were the malignancy-associated markers. The normal adult brain-derived lines showed some differentiated astroglial features and expressed low levels of the malignancy-associated properties. The foetal cultures contained highly differentiated astroglia while the glioma lines showed considerable phenotypic heterogeneity from highly differentiated to undifferentiated. The least differentiated glioma cells exhibited the highest plasminogen activator activities. The density-dependent control of phenotypic expression was also investigated. High affinity GABA uptake, and GFAP in rat C6 glioma cultures, increased with increasing monolayer cell density, events probably mediated by an increase in the formation of cell-cell contacts at confluence. Plasminogen activator activity decreased with increasing cell density.The differences between neoplastic cells and their normal counterparts can usually be described in terms of the repression of specific endogenous genes and the inappropriate expression of others. The observed effect is often the absence of differentiated cell products and the acquisition of properties associated with tumour growth and spread. The objective underlying the present investigation was to determine whether a relationship exists between expression of differentiated properties and malignancy-associated properties in early passage cell cultures derived from anaplastic astrocytomas, normal adult and foetal brain. The successful growth of normal and malignant glial cells provides one of the few model systems with the potential for comparing cytology, biochemistry, immunology and behaviour of malignant and normal cells of similar lineage.Marker properties representing the mature differentiated and malignancy-associated astroglial phenotypes were identified and biological, biochemical or immunological assays used to quantitate their levels of expression. The properties associated with astroglial differentiation are closely concerned with some of the specific functions these cells perform in the brain in vivo and have been identified in glial cells in vitro. These were glial fibrillary acidic protein (GFAP) (Eng et al., 1971),

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Plasminogen activator in cultured Lewis lung carcinoma cells measured by chromogenic substrate assay

Cited by 26 publications

References 14 publications

Biological Activity of Retinoids Correlates with Affinity for Nuclear Receptors but Not for Cytosolic Binding Protein

Biological Activity of Retinoids Correlates with Affinity for Nuclear Receptors but Not for Cytosolic Binding Protein

Activity of interferon α, interleukin 6 and insulin in the regulation of differentiation in A549 alveolar carcinoma cells

Interrelationship between differentiation and malignancy-associated properties in glioma

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