TY - JOUR
T1 - The role of protein kinase C α and ε isozymes in DNA synthesis induced by muscarinic receptors in a glial cell line
AU - Guizzetti, Marina
AU - Wei, Min
AU - Costa, Lucio G.
N1 - Funding Information:
This study was supported in part by grants from NIAAA (AA-08154), NIEHS (ES 08851 and ES 07033), and the Alcohol and Drug Abuse Institute, University of Washington.
PY - 1998/10/23
Y1 - 1998/10/23
N2 - Acetylcholine has been shown to induce proliferation of human astrocytoma cells by activating muscarinic receptors, particularly the m3 subtype. In the present study the role of protein kinase C in DNA synthesis induced by carbachol has been investigated. Carbachol-induced [methyl-3H]thymidine incorporation was inhibited by the protein kinase C inhibitors GF 109203X and staurosporine. However, carbachol-induced DNA synthesis was only partially reduced by protein kinase C down-regulation by phorbol 12-myristate 13-acetate (PMA), and maximal concentrations of carbachol and PMA had an additive effect on [methyl-3H]thymidine incorporation. Exposure for 24 h to maximally effective concentrations of carbachol did not induce down-regulation of protein kinase C α, and caused a small but significant down-regulation of protein kinase C ε; cells exposed for 24 h to carbachol were still able to respond with protein kinase C translocation to PMA stimulation. Carbachol caused a significant increase of phorbol ester binding, but did not stimulate protein kinase C α translocation, while it caused a short-lasting translocation of protein kinase C ε; however, protein kinase C ε translocation was not correlated with the time-course of carbachol-induced increase in [methyl-3H]thymidine incorporation. On the other hand, the time-course of translocation/down-regulation of protein kinase C α and protein kinase C ε induced by PMA was in good correlation with the time-course of PMA-induced [methyl-3H]thymidine incorporation. These results suggest that protein kinase C α may not be involved in DNA synthesis induced by muscarinic receptors stimulation in 132-1N1 astrocytoma cells, while protein kinase C ε appears to play a role in the initial exit from G0/G1 phase, though it cannot be considered the major determinant for sustained proliferation. Copyright (C) 1998 Elsevier Science B.V.
AB - Acetylcholine has been shown to induce proliferation of human astrocytoma cells by activating muscarinic receptors, particularly the m3 subtype. In the present study the role of protein kinase C in DNA synthesis induced by carbachol has been investigated. Carbachol-induced [methyl-3H]thymidine incorporation was inhibited by the protein kinase C inhibitors GF 109203X and staurosporine. However, carbachol-induced DNA synthesis was only partially reduced by protein kinase C down-regulation by phorbol 12-myristate 13-acetate (PMA), and maximal concentrations of carbachol and PMA had an additive effect on [methyl-3H]thymidine incorporation. Exposure for 24 h to maximally effective concentrations of carbachol did not induce down-regulation of protein kinase C α, and caused a small but significant down-regulation of protein kinase C ε; cells exposed for 24 h to carbachol were still able to respond with protein kinase C translocation to PMA stimulation. Carbachol caused a significant increase of phorbol ester binding, but did not stimulate protein kinase C α translocation, while it caused a short-lasting translocation of protein kinase C ε; however, protein kinase C ε translocation was not correlated with the time-course of carbachol-induced increase in [methyl-3H]thymidine incorporation. On the other hand, the time-course of translocation/down-regulation of protein kinase C α and protein kinase C ε induced by PMA was in good correlation with the time-course of PMA-induced [methyl-3H]thymidine incorporation. These results suggest that protein kinase C α may not be involved in DNA synthesis induced by muscarinic receptors stimulation in 132-1N1 astrocytoma cells, while protein kinase C ε appears to play a role in the initial exit from G0/G1 phase, though it cannot be considered the major determinant for sustained proliferation. Copyright (C) 1998 Elsevier Science B.V.
KW - Astrocytoma cell
KW - Cell proliferation
KW - Muscarinic receptor
KW - Protein kinase C
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U2 - 10.1016/S0014-2999(98)00620-7
DO - 10.1016/S0014-2999(98)00620-7
M3 - Article
C2 - 9832394
AN - SCOPUS:0032561168
SN - 0014-2999
VL - 359
SP - 223
EP - 233
JO - European Journal of Pharmacology
JF - European Journal of Pharmacology
IS - 2-3
ER -