Accumulation of inositol phosphates in low-passage human meningioma cells following treatment with epidermal growth factor

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✓ In order to elucidate some of the signal transduction processes in human meningioma cells, the authors studied the effect of epidermal growth factor (EGF) and bromocriptine on inositol phospholipid hydrolysis, using low-passage human meningioma cells in culture. Epidermal growth factor is a well-studied mitogenic factor for meningioma cells, whereas bromocriptine is known to have an inhibitory effect on meningioma cell proliferation. The addition of EGF to meningioma cells caused stimulation of inositol phosphate accumulation in a dose-dependent manner at 60 minutes posttreatment, with the maximum effect (120% to 167% of control) achieved at a concentration of 10 ng/ml. Extraction of separate inositol phosphates revealed that inositol monophosphate (IP1) and inositol bisphosphate (IP2), but not inositol trisphosphate (IP3), accounted for the increase at 60 minutes. Kinetic analysis of EGF-stimulated inositol phospholipid hydrolysis showed that a sharp and transient increase in IP3 from 5 to 12 minutes post-EGF and a transient but more gradual increase in IP2 from 2 to 12 minutes post-EGF were followed by a gradual and steady increase in IP1, which was significantly greater than control after 5 minutes. On the other hand, long-term studies showed a down-regulation of inositol phosphate accumulation (a 64% decrease vs. control) after 7 days of treatment with EGF (10 ng/ml). Bromocriptine (5 µM) exhibited no significant effect on inositol phosphate accumulation at 60 minutes in four of five meningiomas studied. However, of two meningiomas studied with bromocriptine in combination with EGF, both showed a significant additive increase in inositol phosphate accumulation compared to those treated with EGF alone. The results suggest a close involvement of inositol phospholipid turnover in human meningioma cells in response to mitogenic stimulation by EGF.

Article Information

Address reprint requests to: Tomoki Todo, M.D., Department of Neurosurgery, International Medical Center of Japan, 1-21-1 Toyama, Shinjuku-ku, Tokyo 162, Japan.

© AANS, except where prohibited by US copyright law.

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Figures

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    Graphs showing the short-term effect of epidermal growth factor (EGF) on inositol phosphate accumulation in human meningioma cells. Statistical significance: * = p < 0.05 and ** = p < 0.01 versus control value of the corresponding inositol phosphate fraction; vertical bars = standard deviation. Left: Total inositol phosphate formation in response to different doses of EGF at 60 minutes posttreatment was measured by eluting inositol mono-, bis-, and trisphosphate (IP1, IP2, and IP3) together. Results are expressed as a percentage of the control value. Right: Results of studies in which IP1, IP2, and IP3 were extracted separately 60 minutes after addition of EGF (10 ng/ml). The amount of each inositol phosphate fraction is expressed as a percentage of phospholipids.

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    Graphs showing time course of inositol phosphate formation in the absence (dotted lines) and presence (solid lines) of epidermal growth factor (10 ng/ml) in human meningioma cells. Inositol mono-, bis-, and trisphosphate (IP1, IP2, and IP3) were extracted separately at each time point, and the amounts are expressed as a percentage of phospholipids. Statistical significance: * = p < 0.05, ** = p < 0.02, and *** = p < 0.01 versus control value of the corresponding inositol phosphate fraction at the same time point: vertical bars = standard deviation of the mean of three determinations.

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    Graph showing the effect of bromocriptine on inositol phosphate accumulation in human meningioma cells. Inositol mono-, bis-, and triphosphate (IP1, IP2, and IP3) were extracted separately after a 60-minute treatment with epidermal growth factor (EGF, 10 ng/ml), bromocriptine (BC, 5 µM), or both. Results are expressed as a percentage of phospholipids. Statistical significance: * = p < 0.05, ** = p < 0.02, and *** = p < 0.01 versus control, or EGF and bromocriptine together versus EGF or bromocriptine alone; vertical bars = standard deviation.

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    Graph showing the effects of epidermal growth factor (EGF), bromocriptine (BC), and their combination on the proliferation of human meningioma cells, using the same meningioma tissue studied for bromocriptine-induced inositol phosphate accumulation (shown in Fig. 3). Cell numbers are determined after a 7-day treatment with EGF (10 ng/ml), bromocriptine (5 µM), or both together. Statistical significance: * = p < 0.05, ** = p < 0.01, and *** = p < 0.001 versus control, or EGF and bromocriptine together versus EGF or bromocriptine alone; vertical bars = standard deviation.

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