17β-Estradiol enhances heparin-binding epidermal growth factor-like growth factor production in human keratinocytes

Abstract

Heparin-binding epidermal growth factor-like growth factor (HB-EGF) enhances reepithelialization in wounds. Estrogen is known to promote cutaneous wound repair. We examined the in vitro effects of 17β-estradiol (E2) on HB-EGF production by human keratinocytes. E2 or membrane-impermeable BSA-conjugated E2 (E2-BSA) increased HB-EGF secretion, mRNA level, and promoter activity in keratinocytes. E2 or E2-BSA enhanced in vitro wound closure in keratinocytes, and the closure was suppressed by anti-HB-EGF antibody. Activator protein-1 (AP-1) and specificity protein 1 (Sp1) sites on HB-EGF promoter were responsible for the E2- or E2-BSA-induced transactivation. Antisense oligonucleotides against c-Fos, c-Jun, and Sp1 blocked E2- or E2-BSA-induced HB-EGF transactivation. E2 or E2-BSA enhanced DNA binding and transcriptional activity of AP-1 and generated c-Fos/c-Jun heterodimers by inducing c-Fos expression. E2 or E2-BSA enhanced DNA binding and transcriptional activity of Sp1 in parallel with the enhancement of Sp1 phosphorylation. These effects of E2 or E2-BSA were not blocked by the nuclear estrogen receptor antagonist ICI-182,780 or anti-estrogen receptor-α or -β antibodies but were blocked by inhibitors of G protein, phosphatidylinositol-specific PLC, PKC-α, and MEK1. These results suggest that E2 or E2-BSA may enhance HB-EGF production via activation of AP-1 and Sp1. These effects of E2 or E2-BSA may be dependent on membrane G protein-coupled receptors different from nuclear estrogen receptors and on the receptor-mediated activities of phosphatidylinositol-specific PLC, PKC-α, and MEK1. E2 may enhance wound reepithelialization by promoting HB-EGF production in keratinocytes.

Effect of Blood Collection Tube Types on the Measurement of Human Epidermal Growth Factor

“…In previous studies, it had been shown that adult male plasma/serum EGF level was two-fold higher than female in mice, and testosterone treatment of female mice doubled serum EGF level [29] and that EGF level in tear fluid was higher in male compared to female patients; [30] contrary to these reports, we measured serum EGF levels significantly higher in female compared to male volunteers (p ¼ 0.014). Our data regarding the higher serum EGF levels observed for females could be explained by the roles of some hormones reported previously as for progesterone up-regulating EGF expression [31] and, as for 17-beta estradiol enhancing heparin binding EGFlike growth factor production in human keratinocytes [32] and as well as with increasing EGF levels towards term. [33] Using the ELISA system we developed for measurement of hEGF, hEGF levels were below detectable limits in all plasma samples of all volunteers who participated in this study.…”

c‐jun‐NH2JNK mediates invasive potential and EGFR activation by regulating the expression of HB‐EGF in a urokinase‐stimulated pathway

“…This activity was strongly affected by the dominant negative form of JNK and only minimally affected by the ERK1,2 pathway [Benasciutti et al, 2004]. In addition, studies on human keratinocytes have shown that estradiol‐dependent wound re‐epithelialization depends directly on HB‐EGF transcription enhancement which in turn, also depends on AP‐1 and Sp1 [Kanda and Watanabe, 2005]. These data strongly suggest that in our system uPA and HB‐EGF could share a similar transcriptional mechanism with JNK playing a central role.…”

Histamine enhances the production of human β-defensin-2 in human keratinocytes

“…Antisense oligonucleotides against NF-B p50 and p65, STAT1, c-Jun and c-Fos, and control scrambled oligonucleotides were synthesized by phosphoroamidite chemistry as described previously (19,22). Keratinocytes were transfected with the indicated oligonucleotides (0.2 M each) premixed with Fugene 6 in KGM for 6 h. The concentration of antisense oligonucleotides (0.2 M) was selected because this concentration did not reduce the viability of keratinocytes (Ͼ94% viable) and selectively reduced the levels of respective proteins in our previous study (22). The cells were treated with high-calcium KBM for 48 h and then incubated with 1 M histamine and/or 10 ng/ml IFN-␥ or TNF-␣.…”