TY - JOUR
T1 - Metalloproteinase-mediated, context-dependent function of amphiregulin and hb-egf in human keratinocytes and skin
AU - Stoll, Stefan W.
AU - Johnson, Jessica L.
AU - Bhasin, Ajay
AU - Johnston, Andrew
AU - Gudjonsson, Johann E.
AU - Rittié, Laure
AU - Elder, James T.
N1 - Funding Information:
We thank Suzan Rehbine for her excellent technical assistance with skin biopsies and Dr James Rheinwald (Harvard Medical School, Boston, MA) for providing N-TERT-2G keratinocytes. This work was in part supported by the National Institute for Arthritis, Musculoskeletal and Skin Disease (NIAMS), National Institutes of Health (NIH award K01 AR050462 and R03 AR049420 to SWS and R01 AR 052889 to JTE) and by a Chesebrough Pond's Lever Brothers Dermatology Foundation Research Career Development Award (SWS). JTE is supported by the Ann Arbor Veterans Affairs Hospital.
PY - 2010/1/1
Y1 - 2010/1/1
N2 - Human keratinocytes (KCs) express multiple EGF receptor (EGFR) ligands; however, their functions in specific cellular contexts remain largely undefined. To address this issue, first we measured mRNA and protein levels for multiple EGFR ligands in KCs and skin. Amphiregulin (AREG) was by far the most abundant EGFR ligand in cultured KCs, with >19 times more mRNA and >7.5 times more shed protein than any other family member. EGFR ligand expression in normal skin was low (<8% of RPLP0/36B4); however, HB-EGF and AREG mRNAs were strongly induced in human skin organ culture. KC migration in scratch wound assays was highly metalloproteinase (MP)-and EGFR dependent, and was markedly inhibited by EGFR ligand antibodies. However, lentivirus-mediated expression of soluble HB-EGF, but not soluble AREG, strongly enhanced KC migration, even in the presence of MP inhibitors. Lysophosphatidic acid (LPA)-induced ERK phosphorylation was also strongly EGFR and MP dependent and markedly inhibited by neutralization of HB-EGF. In contrast, autocrine KC proliferation and ERK phosphorylation were selectively blocked by neutralization of AREG. These data show that distinct EGFR ligands stimulate KC behavior in different cellular contexts, and in an MP-dependent fashion.
AB - Human keratinocytes (KCs) express multiple EGF receptor (EGFR) ligands; however, their functions in specific cellular contexts remain largely undefined. To address this issue, first we measured mRNA and protein levels for multiple EGFR ligands in KCs and skin. Amphiregulin (AREG) was by far the most abundant EGFR ligand in cultured KCs, with >19 times more mRNA and >7.5 times more shed protein than any other family member. EGFR ligand expression in normal skin was low (<8% of RPLP0/36B4); however, HB-EGF and AREG mRNAs were strongly induced in human skin organ culture. KC migration in scratch wound assays was highly metalloproteinase (MP)-and EGFR dependent, and was markedly inhibited by EGFR ligand antibodies. However, lentivirus-mediated expression of soluble HB-EGF, but not soluble AREG, strongly enhanced KC migration, even in the presence of MP inhibitors. Lysophosphatidic acid (LPA)-induced ERK phosphorylation was also strongly EGFR and MP dependent and markedly inhibited by neutralization of HB-EGF. In contrast, autocrine KC proliferation and ERK phosphorylation were selectively blocked by neutralization of AREG. These data show that distinct EGFR ligands stimulate KC behavior in different cellular contexts, and in an MP-dependent fashion.
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U2 - 10.1038/jid.2009.211
DO - 10.1038/jid.2009.211
M3 - Article
C2 - 19609315
AN - SCOPUS:72049110515
SN - 0022-202X
VL - 130
SP - 295
EP - 304
JO - Journal of Investigative Dermatology
JF - Journal of Investigative Dermatology
IS - 1
ER -