Androgen Receptor is Overexpressed in boys with Severe
Hypospadias and ZEB1 Regulates Androgen Receptor
Expression in Human Foreskin Cells. ZEB1 is overexpressed in
patients with severe hypospadias. We examined the interaction
between ZEB1 and the androgen receptor (AR) in vitro and the
expression of AR in boys with hypospadias.
Hs68 cells, a fibroblast cell line derived from neonatal human
foreskin, were exposed to 0, 10, and 100nM of estrogen, after
which the cellular localization of ZEB1 and AR was assessed
using immunocytochemistry. To determine if ZEB1 interacted
with the AR gene, chromatin immunoprecipitation (ChIP) was
performed using ZEB1 antibody and PCR for AR. Secondly, AR
expression was quantified using real-time PCR and Western blot
in normal subjects (n = 32), and subjects with mild (n = 16) and
severe hypospadias (n = 16).
ZEB1 and AR co-localize to the nucleus. Estrogen upregulated
ZEB1 and AR expression. ChIP demonstrated that ZEB1 binds
to an E-box sequence in the AR gene promoter. AR expression is
higher in subjects with severe hypospadias than those with mild
hypospadias and control subjects (p <0.05).
ZEB1 physically interacts with AR in human foreskin cells.
AR is overexpressed in patients with severe hypospadias.
Environmental estrogenic compounds may increase the risk of
hypospadias be facilitating the interaction between ZEB1 and AR.