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Involvement of human beta-defensin-2 in intracellular signaling: in vitro study
D Lytvyn1, E Zhuravel, N Markeeva
1R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology NAS of Ukraine, Kyiv, Ukraine.
Aim:
To analyze involvement of human beta-defensin-2 (hBD-2) in intracellular signaling in vitro.
Materials And Methods:
A431cells were cultured in the presence of 1 microg/ml of recombinant hBD-2 and/or 10 ng/ml EGF. For evaluation of expression of mRNAs for p70S6 kinase, isoforms alpha and beta, RT-PCR analysis was applied. Expression and activity of p70S6K, phosphorylation of PDK1, ERK, JNK, p38 kinases and EGF receptor (EGFR) was evaluated using Western blot analysis.
Results:
30 min incubation of A431 cells with 1 mug/ml of hBD-2 didn't influence autophosphorylation level of EGFR, but resulted in activation of p70S6K, 12 h treatment - in prominently increased level of mRNA for alpha and beta-isoforms of p70S6 kinase, whilst 24 h treatment - in elevation of p70S6K synthesis on protein level. Up-stream kinase phosphorylating p70S6K, PDK1, is also phosporylated upon influence of exogenous hBD-2 in vitro.
Conclusion:
Our data point on the involvement of PDK1-p70S6K pathway in mediation of action of hBD-2 in A431 cells.
