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Biomedical subjects

M A Fang

Publications and source records attributed to M A Fang.

9 recordsLinked to original sources

Epidermal growth factor induces Egr-1 messenger RNA and protein in mouse osteoblastic cells.

The nuclear signaling events activated when epidermal growth factor (EGF) interacts with osteoblasts to produce effects on growth and differentiation are not clearly understood, and may include induction of immediate early genes such as Egr-1, a zinc finger transcription factor. In the present study, Northern analyses were performed to define the effects of EGF on the expression of Egr-1 mRNA in MC3T3-E1 mouse osteoblastic cells. Following treatment of quiescent, subconfluent MC3T3-E1 cells with 0.1-100 ng/ml EGF for various periods, maximal induction of Egr-1 mRNA occurred when cells were treated for 30-60 minutes with 1-10 ng/ml EGF. Inhibition of protein kinase C activity by pretreatment with 1 microM chelerythrine chloride or by prolonged stimulation with 50 ng/ml tetradecanoyl phorbol acetate (TPA) partially diminished the induction of Egr-1 by EGF. Using an immunohistochemical approach, 10 ng/ml EGF was observed to induce Egr-1 protein within 30-60 minutes and this induction was localized to the nucleus. These observations indicate that EGF induces Egr-1 mRNA and protein via protein kinase C and other signaling pathways, and that Egr-1 may be part of the regulatory network mediating the actions of EGF on growth and differentiation of osteoblasts.

Animals

The effects of prostaglandin E2, parathyroid hormone, and epidermal growth factor on mitogenesis, signaling, and primary response genes in UMR 106-01 osteoblast-like cells.

Prostaglandin E2 (PGE2), PTH, and epidermal growth factor (EGF) are potent regulators of osteoblast proliferation. In UMR 106-01 rat osteosarcoma cells with osteoblast-like features, PGE2 and PTH inhibit, while EGF stimulates, mitogenesis. Both PGE2 and PTH increase intracellular cAMP levels, cytosolic calcium, and inositol phosphate turnover. In a variety of cell types, EGF mediates its effects in part via activation of receptor protein-tyrosine kinase and other protein kinases, such as protein kinase-C. The nuclear mechanisms of PGE2, PTH, and EGF regulation of osteoblast proliferation are unknown. Accordingly, we have examined the effects of these agents on mitogenesis, second messenger generation, and primary response genes, which may link second messenger activation to subsequent alterations in gene expression. Northern blot analysis of mRNA from UMR 106-01 cells treated for 3 h with 2 microM PGE2, 10 nM PTH, or 10 ng/ml EGF in the presence of cycloheximide demonstrated that all three agents induced the expression of c-fos and c-jun mRNA. In contrast, only EGF stimulated cellular proliferation and induced Egr-1 mRNA. Also, unlike PGE2 and PTH, EGF did not increase intracellular cAMP levels. c-fos mRNA was induced by treatment with 50 ng/ml tetradecanoyl phorbol acetate or by 40 ng/ml forskolin, while induction of Egr-1 mRNA was stimulated by treatment with tetradecanoyl phorbol acetate, but not forskolin. Thus, EGF signal transduction differs from that of PGE2 and PTH in UMR 106-01 osteoblast-like cells, in that EGF does not stimulate the protein kinase-A second messenger system, but causes activation of Egr-1, a primary response gene that may play a role in the mitogenic effect of EGF.

Animals

Effects of interleukin-6 on cellular function in UMR-106-01 osteoblastlike cells.

High levels of interleukin-6 (IL-6) have been detected in synovial fluid from patients with inflammatory arthropathies associated with local bone resorption, suggesting a role for IL-6 as a local regulator of bone resorption and remodeling. In the present study we examined the effects of IL-6 on [3H]thymidine ([3H]TdR) incorporation, collagen synthesis, and alkaline phosphatase activity in UMR-106-01 rat osteoblastic osteosarcoma cells. IL-6 stimulated a dose-dependent increase in [3H]TdR incorporation that was maximal at 1000 U/ml (-147% of basal, p less than 0.005) in osteoblastlike cells that were in a logarithmic phase of growth. The increase in [3H]TdR incorporation was maximal between 12 and 24 h and was neutralized by pretreatment with the polyclonal rabbit antibody to IL-6. IL-6 also increased cell number and the secretion of prostaglandin E2 in UMR-106-01 cells in logarithmic growth phase. The stimulation of [3H]TdR incorporation and release of PGE2 into the culture medium by IL-6 was inhibited by indomethacin. A 24 h exposure of the osteoblastlike cells to 1000 U/ml of IL-6 reduced [3H]proline incorporation into collagenase-digestible (CDP) protein to 73% of control values (p less than 0.01). Noncollagen protein (NCP) synthesis was inhibited to 80% of control values (p less than 0.01) by 1000 U/ml of IL-6. The inhibitory effect was relatively greater on CDP than on NCP and consequently resulted in a decrease in the percentage of collagen synthesis. Alkaline phosphatase activity was not altered in these cells after a 24 h exposure to 1-1000 U/ml of IL-6.(ABSTRACT TRUNCATED AT 250 WORDS)

Alkaline Phosphatase

Effects of nicotine on cellular function in UMR 106-01 osteoblast-like cells.

We examined the effect of nicotine on cellular proliferation, as measured by [3H]thymidine (TdR) incorporation and cell count, and on alkaline phosphatase activity in UMR 106-01 rat osteoblastic osteosarcoma cells. The cells were cultured with varying concentrations of nicotine in serum-free medium for 2 to 72 hours. Nicotine produced a dose-dependent suppression of TdR incorporation, with maximum suppression seen at 10 mM (7% of control); the EC50 for suppression of TdR incorporation was 10 microM. 1 microM nicotine decreased cell number by 20% to 30%. The time course of the effect of 100 microM nicotine on DNA synthesis was measured by TdR incorporation. TdR uptake was measured at 2, 4, 6, 24, 48, and 72 hours. After the addition of nicotine, the following biphasic response in TdR incorporation was observed: a 15% decrease at 2 hours, recovery to near control value at 6 hours, a 27% decrease by 24 hours, and a maximum decrease of 88% by 48 hours. Over a dose range of 1 nM to 10 mM, nicotine produced a dose-dependent increase in alkaline phosphatase activity with maximum stimulation seen at 1 microM (189% of control). We conclude that nicotine suppresses cellular proliferation and stimulates alkaline phosphatase activity in UMR 106-01 osteoblast-like cells. These results may be of significance in the development of osteoporosis and alveolar bone loss associated with the use of tobacco.

Alkaline Phosphatase

Subacute perimyositis.

We describe a patient who developed a benign myopathy with features suggestive of subacute perimysial inflammation. There was no blood eosinophilia or infectious agent identified. Creatine phosphokinase was not significantly elevated. Electromyography showed a myopathic process. Muscle biopsy showed predominantly subacute perimysial inflammation with rare eosinophils. The patient recovered completely after therapy with glucocorticoids and a nonsteroidal antiinflammatory drug. The features of our case were compared to known causes of myositis with or without eosinophils. It may be part of the spectrum of relapsing eosinophilic perimyositis or may be a condition previously unreported.

Acute Disease

Polyarteritis nodosa of the epididymis in a patient with Whipple's disease.

The case of a 55-year-old white male who developed necrotizing arteritis localized to the superior pole of the epididymis is presented. He had a history of Whipple's disease and euthyroid Graves' disease. Histopathologic section of an extratesticular mass showed a necrotizing vasculitis with giant cells; periodic acid Schiff stain for Whipple "bacilli" was negative. The combination of rare diseases in our patient suggests the possibility of a common infectious or immune etiology, perhaps mediated via circulating immune complexes.

Epididymis

Technetium-99m MDP demonstration of heterotopic ossification in fibrodysplasia ossificans progressiva.

The extensive heterotopic bone formation in patients with fibrodysplasia ossificans progressiva (FOP) has been documented previously with radiographs. A case in which a Tc-99m MDP bone scan showed increased uptake at sites well before ossification could be documented radiographically is described. This finding suggests that bone scans would likely be useful to monitor the extent of involvement with FOP and to detect areas of new activity.

Child