SMAD2 and SMAD7 involvement in the post-translational regulation of Muc4 via the transforming growth factor-β and interferon-γ pathways in rat mammary epithelial cells

Muc4/sialomucin complex (SMC) is a heterodimeric glycoprotein complex derived from a single gene that is post-translationally processed into mucin (ASGP-1) and transmembrane (ASGP-2) subunits. Muc4/SMC is tightly regulated in the rat mammary gland, low in the virgin, increased during pregnancy and lactation, and overexpressed in some aggressive mammary tumors. Investigations of primary rat mammary epithelial cells (MEC) have shown that Muc4/SMC expression is post-translationally regulated through inhibition of Muc4/SMC precursor processing by transforming growth factor-beta (TGF-beta). Localization studies suggest that TGF-beta inhibition of Muc4/SMC expression is mediated through SMAD2, a TGF-beta effector that, when activated, functions as a transcription factor. SMAD2 antisense oligonucleotide blocks the inhibition of Muc4/SMC expression by TGF-beta. The TGF-beta effect on Muc4/SMC expression is repressed by interferon-gamma (IFN-gamma). IFN-gamma treatment of MEC activates and relocalizes signal transducer and activator of transcription-1 (STAT-1) to induce an inhibitor SMAD, SMAD7. SMAD7 antisense oligonucleotide prevents IFN-gamma from blocking the TGF-beta inhibition of Muc4/SMC expression. These results suggest that TGF-beta regulates Muc4/SMC expression via the SMAD pathway by a transcriptional effect on a protein in the Muc4/SMC processing step, possibly the protease that cleaves the precursor.