Naturally occurring phenolic acids modulate TPA-induced activation of EGFR, AP-1, and STATs in mouse epidermis.

Epidermal growth factor receptor (EGFR) plays an important role in epithelial carcinogenesis and appears to be involved in STATs activation. In this study we investigated the possible interference of naturally occurring phenolic acids with EGFR, activator protein-1 (AP-1), and signal transducers and activators of transcription (STATs) pathways activated by topical application of tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) in Balb/c mice epidermis. Pretreatment with tannic or chlorogenic acid resulted in a significant decrease in the phosphorylation of EGFR Y-1068 and Y-1173 tyrosine residues, which was accompanied by reduced activation of AP-1. Tannic acid decreased also the c-Jun AP-1 subunit level and binding to TPA response element (TRE) (3- and 2-fold in comparison with TPA-treated group respectively). Simultaneous reduction of JNK activity might be responsible for reduced activation of AP-1. In contrast to these more complex phenolics, protocatechuic acid increased the activity of JNK and was also the most efficient inhibitor of STATs activation. These results indicate that naturally occurring phenolic acids, by decreasing EGFR, AP-1, and STATs activation, may modulate other elements both upstream and downstream in these pathways and thus inhibit the tumor development. Although more complex phenolics affect mainly the EGFR/AP-1 pathway, STATs seem to be the most important targets for simple compounds, such as protocatechuic acid.

Correlation between EGFR Y1068 tyrosine phosphorylation and AP-1 activation by tumor promoter 12-O-tetradecanoylphorbol-13-acetate in mouse skin.

The mouse skin carcinogenesis is unique model for our understating of molecular events leading to tumor development. The tumor promoter, 12-O-tetradecanoylphorbol-13-acetate (TPA) activates a variety of signaling pathways, including MAPK/AP-1. In this study, we examined the time course of EGFR phosphorylation and AP-1 activation in mouse epidermis after topical application of a single 10 nmol dose of TPA. Remarkable differences in the phosphorylation kinetics of EGFR tyrosine residues were observed. While the maximal level of Y1068 tyrosine phosphorylation occurred 4h after TPA treatment, the Y1173 residue phosphorylation was initially down-regulated, and reached the highest level after 24 h. Phosphorylation of Y1068 tyrosine was correlated with AP-1 activation and c-Jun N-terminal kinase (JNK) activity. These results indicate that the stimulation of AP-1 in mouse epidermis by TPA may be the effect of EGFR activation, but not all tyrosine residues forming its catalytic center are equally involved in this process.

[RasGRP proteins--Ras-activating factors]. / Bialka RasGRP--czynniki aktywujace Ras.

The Ras proteins, members of small GTP-binding protein family, are regulated through the exchange of GTP/GDP nucleotide. The activity of the Ras proteins is controlled by guanine nucleotide exchange factors (GEFs) and GTP-ase activating proteins (GAPs), which activate and inactivate G proteins respectively. Beside other, well known Ras-activating GEFs, the new class of such factors was recently described. RasGRP family, known also as CalDAG-GEF, consists of four members. C1 domain, allows them to bind diacylglycerol as well as DAG-analogs like phorbol esters. Binding of the ligand leads to activation of RasGRPs and in consequence to the activation of Ras and Rap proteins by the exchange of bounded guanine nucleotides. The signal transmitted by RasGRP is terminated as a result of DAG phosphorylation catalyzed by diacylglycerol kinase (DGK). Location of RasGRP proteins on the crossing of signaling cascades and broad tissue expression pattern involve them in many events essential for the cell function. RasGRP proteins play roles in such phenomena as: T cells maturation and functioning, B cells response, platelet aggregation, mast cells activity regulation, transformation and many other. In this review, structure and function of RasGRP proteins, as well as their role in neoplastic transformation are described.