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  • Title: The role of protein kinase C alpha and epsilon isozymes in DNA synthesis induced by muscarinic receptors in a glial cell line.
    Author: Guizzetti M, Wei M, Costa LG.
    Journal: Eur J Pharmacol; 1998 Oct 23; 359(2-3):223-33. PubMed ID: 9832394.
    Abstract:
    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 alpha, and caused a small but significant down-regulation of protein kinase C epsilon; 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 alpha translocation, while it caused a short-lasting translocation of protein kinase C epsilon; however, protein kinase C epsilon 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 alpha and protein kinase C epsilon 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 alpha may not be involved in DNA synthesis induced by muscarinic receptors stimulation in 132-1N1 astrocytoma cells, while protein kinase C epsilon appears to play a role in the initial exit from G0/G1 phase, though it cannot be considered the major determinant for sustained proliferation.
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