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Rapid responses to dexamethasone in the 16HBE14o‐ human bronchial epithelial cell line
Author(s) -
Hynes Darina,
Harvey Brian J
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.5.a1440-c
Subject(s) - protein kinase c , kinase , p38 mitogen activated protein kinases , mapk/erk pathway , microbiology and biotechnology , protein kinase a , mitogen activated protein kinase , signal transduction , biology , chemistry
Our previous studies demonstrated a rapid anti‐secretory effect of dexamethasone (DEXA) treatment in the 16HBE14o‐ human bronchial epithelial cell line and identified protein kinase signalling pathways mediating this response. Expression of protein kinases was investigated using RT‐PCR and Western blotting while kinase activation was determined using specific phospho‐antibodies. Firstly, we examined the basal expression of protein kinase C isoforms (PKCα, PKCδ, and PKCε) and of protein kinase D (PKD) a known downstream target of PKC. In addition, we determined the expression and activation of ERK 1/2, p38 and SAPK/JNK kinases. The 16HBE14o‐ cell line was found to express the full complement of the investigated kinases. The relative expression levels of PKC isoforms was PKCα > PKCε > PKCδ. DEXA (1 nM) treatment produced a rapid and transient activation of PKCα (2–5 min) and PKD (5–10 min). The activation of PKD was blocked by pre‐treatment with 1nM Bisindolylmaleimide I, a general PKC inhibitor, suggesting a PKC‐dependent activation of PKD. DEXA also produced a later activation (5–15 min) of SAPK/JNK, p38 and ERK 1/2. Since the activation of ERK 1/2 was dependent on Protein Kinase A (PKA), we determined the expression of adenylyl cyclase (AC) isoforms, the upstream regulator of PKA. 16HBE14o‐ expressed four of the ten known AC isoforms (III, IV, VI and VII). Our results show that DEXA rapidly activates several protein kinases in the 16HBE14o‐ cell line. The role of these kinases in mediating rapid responses to glucocorticoids in lung epithelia is discussed. Funded by a HEA‐PRTLI Programme Grant.