title: Loss of TGF-β adaptor β2SP activates notch signaling and SOX9 expression in esophageal adenocarcinoma.
abstract: TGF-β and Notch signaling pathways play important roles in regulating self-renewal of stem cells and gastrointestinal carcinogenesis. Loss of TGF-β signaling components activates Notch signaling in esophageal adenocarcinoma, but the basis for this effect has been unclear. Here we report that loss of TGF-β adapter β2SP (SPNB2) activates Notch signaling and its target SOX9 in primary fibroblasts or esophageal adenocarcinoma cells. Expression of the stem cell marker SOX9 was markedly higher in esophageal adenocarcinoma tumor tissues than normal tissues, and its higher nuclear staining in tumors correlated with poorer survival and lymph node invasion in esophageal adenocarcinoma patients. Downregulation of β2SP by lentivirus short hairpin RNA increased SOX9 transcription and expression, enhancing nuclear localization for both active Notch1 (intracellular Notch1, ICN1) and SOX9. In contrast, reintroduction into esophageal adenocarcinoma cells of β2SP and a dominant-negative mutant of the Notch coactivator mastermind-like (dnMAN) decreased SOX9 promoter activity. Tumor sphere formation and invasive capacity in vitro and tumor growth in vivo were increased in β2SP-silenced esophageal adenocarcinoma cells. Conversely, SOX9 silencing rescued the phenotype of esophageal adenocarcinoma cells with loss of β2SP. Interaction between Smad3 and ICN1 via Smad3 MH1 domain was also observed, with loss of β2SP increasing the binding between these proteins, inducing expression of Notch targets SOX9 and C-MYC, and decreasing expression of TGF-β targets p21(CDKN1A), p27 (CDKN1B), and E-cadherin. Taken together, our findings suggest that loss of β2SP switches TGF-β signaling from tumor suppression to tumor promotion by engaging Notch signaling and activating SOX9.
authors.affiliation: Department of Gastroenterology, Hepatology & Nutrition, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA. ssong@mdanderson.org
authors.forename: Shumei
authors.initials: S
authors.lastname: Song
authors.forename: Dipen M
authors.initials: DM
authors.lastname: Maru
authors.forename: Jaffer A
authors.initials: JA
authors.lastname: Ajani
authors.forename: Chia-Hsin
authors.initials: CH
authors.lastname: Chan
authors.forename: Soichiro
authors.initials: S
authors.lastname: Honjo
authors.forename: Hui-Kuan
authors.initials: HK
authors.lastname: Lin
authors.forename: Arlene
authors.initials: A
authors.lastname: Correa
authors.forename: Wayne L
authors.initials: WL
authors.lastname: Hofstetter
authors.forename: Marta
authors.initials: M
authors.lastname: Davila
authors.forename: John
authors.initials: J
authors.lastname: Stroehlein
authors.forename: Lopa
authors.initials: L
authors.lastname: Mishra
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics / Q000378:metabolism* / Q000401:mortality; D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D002352:Carrier Proteins / Q000502:physiology*; D002454:Cell Differentiation; D002465:Cell Movement; D049109:Cell Proliferation; D047369:Chromatin Immunoprecipitation; D004938:Esophageal Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000401:mortality; D005434:Flow Cytometry; D006801:Humans; D047468:Immunoprecipitation; D008207:Lymphatic Metastasis; D051379:Mice; D008819:Mice, Nude; D008840:Microfilament Proteins / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D051881:Receptor, Notch1 / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055758:SOX9 Transcription Factor / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D051900:Smad3 Protein / Q000235:genetics / Q000378:metabolism*; D015996:Survival Rate; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D023041:Xenograft Model Antitumor Assays; D013049:Spectrin
chemical_list: D002352:Carrier Proteins; D008840:Microfilament Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D051881:Receptor, Notch1; D055758:SOX9 Transcription Factor; D051900:Smad3 Protein; D016212:Transforming Growth Factor beta; D013049:Spectrin; C497761:NOTCH1 protein, human; C497790:SMAD3 protein, human; C527932:SOX9 protein, human
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