Differential Regulation of Cyclin D1 Expression by Protein Kinase C α and ε Signaling in Intestinal Epithelial Cells

Cellular accumulation of cyclin D1, a key regulator of cell proliferation and tumorigenesis, is subject to tight control. Our previous studies have identified PKC alpha as a negative regulator of cyclin D1 in the intestinal epithelium. However, treatment of non-transformed IEC-18 ileal crypt cells with PKC agonists has a biphasic effect on cyclin D1 expression. Initial PKC alpha-mediated down-regulation is followed by recovery and subsequent accumulation of the cyclin to levels markedly higher than those seen in untreated cells. Using protein overexpression strategies, siRNA, and pharmacological inhibitors, we now demonstrate that the recovery and hyperinduction of cyclin D1 reflect the combined effects of (a) loss of negative signals from PKC alpha due to agonist-induced PKC alpha down-regulation and (b) positive effects of PKC epsilon. PKC epsilon-mediated up-regulation of cyclin D1 requires sustained ERK stimulation and transcriptional activation of the proximal cyclin D1 (CCDN1) promoter, without apparent involvement of changes in protein stability or translation. PKC epsilon also up-regulates cyclin D1 expression in colon cancer cells, through mechanisms that parallel those in IEC-18 cells. Although induction of cyclin D1 by PKC epsilon is dependent on non-canonical NF-kappa B activation, the NF-kappa B site in the proximal promoter is not required. Instead, cyclin D1 promoter activity is regulated by a novel interaction between NF-kappa B and factors that associate with the cyclic AMP-response element adjacent to the NF-kappa B site. The differential effects of PKC alpha and PKC epsilon on cyclin D1 accumulation are likely to contribute to the opposing tumor-suppressive and tumor-promoting activities of these PKC family members in the intestinal epithelium.