title: A NOTCH3-mediated squamous cell differentiation program limits expansion of EMT-competent cells that express the ZEB transcription factors.
abstract: Zinc finger E-box-binding (ZEB) proteins ZEB1 and ZEB2 are transcription factors essential in TGF-β-mediated senescence, epithelial-to-mesenchymal transition (EMT), and cancer stem cell functions. ZEBs are negatively regulated by members of the miR-200 microRNA family, but precisely how tumor cells expressing ZEBs emerge during invasive growth remains unknown. Here, we report that NOTCH3-mediated signaling prevents expansion of a unique subset of ZEB-expressing cells. ZEB expression was associated with the lack of cellular capability of undergoing NOTCH3-mediated squamous differentiation in human esophageal cells. Genetic inhibition of the Notch-mediated transcriptional activity by dominant-negative Mastermind-like 1 (DNMAML1) prevented squamous differentiation and induction of Notch target genes including NOTCH3. Moreover, DNMAML1-enriched EMT-competent cells exhibited robust upregulation of ZEBs, downregulation of the miR-200 family, and enhanced anchorage-independent growth and tumor formation in nude mice. RNA interference experiments suggested the involvement of ZEBs in anchorage-independent colony formation, invasion, and TGF-β-mediated EMT. Invasive growth and impaired squamous differentiation were recapitulated upon Notch inhibition by DNMAML1 in organotypic three-dimensional culture, a form of human tissue engineering. Together, our findings indicate that NOTCH3 is a key factor limiting the expansion of ZEB-expressing cells, providing novel mechanistic insights into the role of Notch signaling in the cell fate regulation and disease progression of esophageal squamous cancers.
authors: Gastroenterology Division, Department of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
authors: Shinya
authors: S
authors: Ohashi
authors: Mitsuteru
authors: M
authors: Natsuizaka
authors: Seiji
authors: S
authors: Naganuma
authors: Shingo
authors: S
authors: Kagawa
authors: Sotai
authors: S
authors: Kimura
authors: Hiroshi
authors: H
authors: Itoh
authors: Ross A
authors: RA
authors: Kalman
authors: Momo
authors: M
authors: Nakagawa
authors: Douglas S
authors: DS
authors: Darling
authors: Devraj
authors: D
authors: Basu
authors: Phyllis A
authors: PA
authors: Gimotty
authors: Andres J
authors: AJ
authors: Klein-Szanto
authors: J Alan
authors: JA
authors: Diehl
authors: Meenhard
authors: M
authors: Herlyn
authors: Hiroshi
authors: H
authors: Nakagawa
mesh_terms: D000818:Animals; D015703:Antigens, CD; D015820:Cadherins / Q000096:biosynthesis / Q000235:genetics; D002294:Carcinoma, Squamous Cell / Q000235:genetics / Q000473:pathology*; D002455:Cell Division; D045744:Cell Line, Tumor; D004268:DNA-Binding Proteins / Q000502:physiology; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics / Q000502:physiology*; D004938:Esophageal Neoplasms / Q000235:genetics / Q000473:pathology*; D015972:Gene Expression Regulation, Neoplastic; D018398:Homeodomain Proteins / Q000502:physiology*; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D035683:MicroRNAs / Q000096:biosynthesis / Q000235:genetics; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D009368:Neoplasm Transplantation; D034622:RNA Interference; D012334:RNA, Neoplasm / Q000096:biosynthesis / Q000235:genetics; D034741:RNA, Small Interfering / Q000494:pharmacology; D000071656:Receptor, Notch3; D051880:Receptors, Notch / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000502:physiology; D012097:Repressor Proteins / Q000502:physiology*; D015398:Signal Transduction; D014157:Transcription Factors / Q000502:physiology*; D014410:Tumor Stem Cell Assay; D000074165:Zinc Finger E-box Binding Homeobox 2; D000071799:Zinc Finger E-box-Binding Homeobox 1
chemical_list: D015703:Antigens, CD; D015820:Cadherins; D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D035683:MicroRNAs; D009363:Neoplasm Proteins; D012334:RNA, Neoplasm; D034741:RNA, Small Interfering; D000071656:Receptor, Notch3; D051880:Receptors, Notch; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; D000074165:Zinc Finger E-box Binding Homeobox 2; D000071799:Zinc Finger E-box-Binding Homeobox 1; C513384:CDH1 protein, human; C430872:MAML1 protein, human; C533896:MIRN200 microRNA, human; C534425:MIR205, human; C499374:NOTCH3 protein, human; C486307:ZEB1 protein, human; C492363:ZEB2 protein, human