Genome-wide analysis of endogenously expressed ZEB2 binding sites reveals inverse correlations between ZEB2 and GalNAc-transferase GALNT3 in human tumors

Abstract

Our observation that ZEB2 negatively regulates a GalNAc-transferase (GALNT3) that is involved in O-glycosylation adds another layer of complexity to the role of ZEB2 in cancer progression and metastasis. Proteins glycosylated by GALNT3 may be exploited as novel diagnostics and/or therapeutic targets.

ETS1 is coexpressed with ZEB2 and mediates ZEB2‐induced epithelial‐mesenchymal transition in human tumors

“…ZEB2 antibody pulled‐down sample displayed more than 20‐fold enrichment compared with control antibody suggesting the presence of ZEB2‐binding sites in ETS1 gene (Figure ). This enrichment was comparable to a previously established ZEB2 target gene, LPCAT1 . In addition, we confirmed ChIP results with luciferase reporter assays in ZEB2 ‐inducible DLD1 and ZEB2‐silenced SNU398 cells.…”

“…E‐cadherin‐positive cell lines are considered as epithelial and vimentin expressing ones can be classified as mesenchymal . According to our results, previous findings, and morphological assessment by others HepG2, Hep3B, Huh7, and PLC/PRF/5 cells were characterized as epithelial, whereas SK‐Hep1, SNU182, and SNU423 cells displayed mesenchymal phenotype . We first determined the transcript levels of ETS1 and homologous transcription factors ZEB1 and ZEB2 in HCC cell lines.…”

“…These results suggest that ZEB2 and ETS1 are more closely associated than ZEB1 and ETS1 are, and prompted us to study whether ETS1 expression is regulated in a ZEB2‐dependent manner in HCC. To this end, we silenced the expression of ZEB2 by shZEB2 lentiviral particles in SNU398 and SK‐HEP‐1 cells, which was previously shown to contain high levels of endogenous ZEB2 transcript and protein . The expression of ETS1 protein decreased more than threefold ( P < 0.001) in shZEB2 clone of SNU398 cells compared with control clone (Figure A).…”

“…Hepatoma cell lines HepG2, Hep3B, PLC/PRF/5, HUH7, and SK‐Hep‐1 were cultured in complete Dulbecco's modified Eagle's medium (DMEM), and SNU182 and SNU423 cells were cultured in complete RPMI1640 media supplemented with 10% fetal bovine serum (FBS) and antibiotics. shZEB2‐SNU398 and nonsilencing control shRNA‐SNU398 (NSC‐SNU398) cells were previously described . shZEB2‐ SK‐HEP‐1 and nonsilencing control shRNA‐SK‐HEP‐1 clones were generated from SK‐HEP‐1 cells as previously described .…”

“…Gene expression analyses were performed as previously described . Briefly, RNA was isolated by using NucleoSpin RNA Kit (Macherey‐Nagel, Düren, Germany) and complementary DNA (cDNA) was synthesized using ProtoScript M‐MuLV Taq RT‐PCR Kit (New England Biolabs, Ipswich, MA) according to instructions of manufacturers.…”