epidermal-growth-factor and diadenosine-tetraphosphate

Diadenosine polyphosphates (diadenosine triphosphate, Ap3A; diadenosine tetraphosphate, Ap4A; diadenosine pentaphosphate, Ap5A; diadenosine hexaphosphate, Ap6A) are potent vasoactive molecules stored and released by platelets. We examined whether these dinucleotides might contribute to the glomerular inflammatory response by stimulating the proliferation of mesangial cells. In cultured rat mesangial cells all four tested dinucleotides (10 to 100 mumol/L) significantly stimulated DNA synthesis as measured by [3H]thymidine uptake at 48 hours (x-fold increase compared with unstimulated control cells: Ap3A, 1.5; Ap4A, 1.8; Ap5A, 1.6; Ap6A, 1.6). In combination with the platelet products platelet-derived growth factor, epidermal growth factor, and serotonin, the dinucleotides synergistically increased DNA synthesis. Dinucleotides by themselves increased cell counts by 23% to 43% at day 2 and augmented mesangial cell growth induced by platelet-derived growth factor, epidermal growth factor, and serotonin. Furthermore, dinucleotides (100 mumol/L) rapidly induced a modest increase in expression of the early growth response gene Egr-1 at 30 minutes (x-fold increase over baseline control: Ap3A, 1.9; Ap4A, 2.8; Ap5A, 2.2; Ap6A, 2.1). We found that extracellular Ap4A was metabolized by mesangial cell ectoenzymes to mononucleotides and adenosine, which also have been shown to be mitogenic for mesangial cells. The combination of Ap4A with mononucleotides or adenosine failed to cause additive stimulation of DNA synthesis in mesangial cells. We conclude that diadenosine polyphosphates stimulate proliferation of cultured mesangial cells and augment mesangial cell growth induced by other mitogens released from platelets. Different molecular mechanisms may be involved in dinucleotide-induced mitogenesis of mesangial cells. Direct effects of dinucleotides on cultured mesangial cells. Direct effects of dinucleotides on cultured mesangial cells appear to play a role because dinucleotides rapidly caused activation of Egr-1.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Analysis of Variance; Animals; Blotting, Northern; Cells, Cultured; Dinucleoside Phosphates; DNA; Epidermal Growth Factor; Glomerular Mesangium; Male; Mitosis; Platelet-Derived Growth Factor; Rats; Rats, Sprague-Dawley; Second Messenger Systems; Serotonin

Diadenosine 5',5'''-P1,P4-tetraphosphate (Ap4A) has been proposed as an intracellular signal for growth. In order to test this hypothesis Ap4A levels were followed in several cell types under various conditions. Quiescent dog thyroid cells in a primary culture were induced to proliferate by addition of a mixture of epidermal growth factor, thyrotropin and foetal calf serum; V79 cells were synchronized by serum depletion then stimulated to proliferate by addition of foetal calf serum. Protein and DNA synthesis increased in both cases, although no significant changes in Ap4A levels per cell could be demonstrated. HeLa D98/AH2 and L929 cells were treated with human recombinant tumour necrosis factor alpha which caused marked cell death. This was measured by a decrease in DNA content and a release into extracellular medium of incorporated radioactive precursor. No concomitant variations in Ap4A concentrations could be observed under these conditions. The data from these various systems do not support the hypothesis that changes in Ap4A levels regulate cellular proliferation.

Topics: Animals; Cattle; Cell Division; Cell Survival; Cells, Cultured; Cricetinae; Cricetulus; Dinucleoside Phosphates; DNA Replication; Epidermal Growth Factor; Fibroblasts; HeLa Cells; Humans; Interferon-gamma; L Cells; Lung; Male; Mice; Recombinant Proteins; Thyroid Gland; Thyrotropin; Tumor Necrosis Factor-alpha