Follistatin antagonizes transforming growth factor-beta3-induced epithelial-mesenchymal transition in vitro: implications for murine palatal development supported by microarray analysis.
Epithelial-mesenchymal transition (EMT) is involved in normal embryonic development as well as in tumor progression and invasiveness. This process is also known to be a crucial step in palatogenesis during fusion of the bi-lateral palatal processes. Disruption of this step results in a cleft palate, which is among the most frequent birth defects in humans. A number of genes and encoded proteins have been shown to play a role in this developmental stage. The central role is attributed to the cytokine transforming growth factor-beta3 (TGF-beta3), which is expressed in the medial edge epithelium (MEE) already before the fusion process. The MEE covers the tips of the growing palatal shelves and eventually undergoes EMT or programmed cell death (apoptosis). TGF-beta3 is described to induce EMT in embryonic palates. With regard to the early expression of this molecule before the fusion process, it is not well understood which mechanisms prevent the TGF-beta3 producing epithelial cells from undergoing differentiation precociously. We used the murine palatal fusion to study the regulation of EMT. Specifically, we analyzed the MEE for the expression of known antagonists of TGF-beta molecules using in situ hybridization and detected the gene coding for Follistatin to be co-expressed with TGF-beta3. Further, we could show that Follistatin directly binds to TGF-beta3 and that it completely blocks TGF-beta3-induced EMT of the normal murine mammary gland (NMuMG) epithelial cell line in vitro. In addition, we analyzed the gene expression profile of NMuMG cells during TGF-beta3-induced EMT by microarray hybridization, detecting strong changes in the expression of apoptosis-regulating genes.
Nogai
Hendrik
H
German Arthritis Research Center, Berlin, Germany.
Rosowski
Mark
M
Grün
Joachim
J
Rietz
Anika
A
Debus
Nils
N
Schmidt
Gül
G
Lauster
Carola
C
Janitz
Michal
M
Vortkamp
Andrea
A
Lauster
Roland
R
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D004847:Epithelial Cells / Q000166:cytology*; D005260:Female; D038681:Follistatin / Q000378:metabolism / Q000502:physiology*; D017403:In Situ Hybridization; D008648:Mesoderm / Q000166:cytology*; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D010159:Palate / Q000166:cytology / Q000196:embryology*; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011485:Protein Binding; D053782:Transforming Growth Factor beta3 / Q000378:metabolism / Q000502:physiology*
D017931:DNA Primers; D038681:Follistatin; D053782:Transforming Growth Factor beta3