Assessment of protein kinase C activity in Y1 adrenal cells.

摘要

The overall purpose of my study was to provide evidence for ACTH signalling through PKC in Y1 adrenal tumor cells and to evaluate the potential ability of mouse PKCϵ R to inhibit PKC activity in this cell line. I first evaluated the usefulness of a PKC-dependent luciferase reporter gene, pAP1(PMA)-luc to measure PKC activity in Y1 cells. My results showed that this reporter plasmid only responded to strong activators and potent inhibitors of PKC. My results also showed that this luciferase reporter system had some degree of PKC specificity. In contrast to the earlier hypotheses, I did not observe an effect of ACTH on luciferase reporter vector nor was luciferase activity inhibited by the expression of mouse PKCϵ R protein. The results from Western blot and immunocytochemistry also demonstrated no detectable effect of PKCϵ R protein on the translocation of PKC holoenzyme.