1125 SILENCING NOTCH-1 AND NOTCH-3 PROMOTES EPITHELIAL-TO-MESENCHYMAL TRANSITION (EMT) AND SELF-RENEWAL POTENTIAL IN HUMAN BLADDER CANCER
Bibliographic record
Abstract
You have accessJournal of UrologyBladder Cancer: Basic Research (III)1 Apr 20131125 SILENCING NOTCH-1 AND NOTCH-3 PROMOTES EPITHELIAL-TO-MESENCHYMAL TRANSITION (EMT) AND SELF-RENEWAL POTENTIAL IN HUMAN BLADDER CANCER Wolfgang Jaeger, Tetsutaro Hayashi, Shannon Awrey, Kilian M. Gust, Thomas Cordonnier, Ladan Fazli, Estelle Li, Na Li, Ralph Buttyan, Theodorus H. Van Der Kwast, Bas W. Van Rhiyn, Joachim W. Thueroff, and Peter C. Black Wolfgang JaegerWolfgang Jaeger Vancouver, Canada More articles by this author , Tetsutaro HayashiTetsutaro Hayashi Vancouver, Canada More articles by this author , Shannon AwreyShannon Awrey Vancouver, Canada More articles by this author , Kilian M. GustKilian M. Gust Frankfurt, Germany More articles by this author , Thomas CordonnierThomas Cordonnier Vancouver, Canada More articles by this author , Ladan FazliLadan Fazli Vancouver, Canada More articles by this author , Estelle LiEstelle Li Vancouver, Canada More articles by this author , Na LiNa Li Vancouver, Canada More articles by this author , Ralph ButtyanRalph Buttyan Vancouver, Canada More articles by this author , Theodorus H. Van Der KwastTheodorus H. Van Der Kwast Toronto, Canada More articles by this author , Bas W. Van RhiynBas W. Van Rhiyn Amsterdam, Netherlands More articles by this author , Joachim W. ThueroffJoachim W. Thueroff Mainz, Germany More articles by this author , and Peter C. BlackPeter C. Black Vancouver, Canada More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.740AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Beside its regulatory functions during development, the Notch pathway plays an eminent role in the tumorigenesis of malignancies. There are four different Notch receptors, which demonstrate context-dependent activity and can work antagonistically to each other. We have previously described a role for Notch-2 in promoting EMT and invasion in bladder cancer (BCA). Our objective here was to elucidate the role of Notch-1 and -3 in EMT and invasion, hypothesizing that both promote the epithelial, less invasive phenotype. METHODS Immunohistochemistry was performed on a tissue microarray (TMA) of 104 tumors from patients with muscle invasive BCA. Notch-1 and -3 expression was assessed in a panel of bladder cancer cell lines by Western blot (WB) and correlated to previously established EMT status (based on E-cadherin and ZEB1) and invasive ability. Notch-1 and -3 were silenced by lentiviral shRNA vector in 3 cell lines with high constitutive Notch-1 and -3 expression (UM-UC1, UM-UC6 and UM-UC15). Protein and mRNA expression of genes relevant to EMT, invasion and stemness were profiled by WB, real-time PCR, immunofluorescence and FACS. Proliferation, migration and invasion were measured by crystal violet, scratch, and Boyden chamber assays, respectively. Self-renewal potential was assessed by clonogenic assay and spheroid formation in MammoCult™ medium. RESULTS High expression of Notch-3 receptor was associated with lower pathological stage (p<0.05) and absence of lymphovascular invasion (p<0.05) in the cystectomy specimens. Notch-1 expression did not correlate with pathologic variables. WB revealed expression of Notch-1 and -3 in epithelial bladder cancer cell lines only. Silencing of Notch-1 or Notch-3 in the selected epithelial cell lines promoted EMT at the mRNA and protein levels. Accordingly, phenotypic changes were seen in the form of enhanced migration and invasion but unaltered proliferation. In addition, colony and spheroid formation was increased after specific Notch-3 silencing. The simultaneous knock down of Notch-1 and -3 showed no additive effect. CONCLUSIONS Our results provide evidence that Notch-1 and Notch-3 are involved in invasion and progression of BCA by maintaining an epithelial phenotype and inhibiting EMT and self-renewal potential. This is the opposite of what we have seen previously with Notch-2. Beside verification of our observations in metastatic xenograft models, further investigation is needed to elucidate the molecular mechanisms of these alterations. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e460 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Wolfgang Jaeger Vancouver, Canada More articles by this author Tetsutaro Hayashi Vancouver, Canada More articles by this author Shannon Awrey Vancouver, Canada More articles by this author Kilian M. Gust Frankfurt, Germany More articles by this author Thomas Cordonnier Vancouver, Canada More articles by this author Ladan Fazli Vancouver, Canada More articles by this author Estelle Li Vancouver, Canada More articles by this author Na Li Vancouver, Canada More articles by this author Ralph Buttyan Vancouver, Canada More articles by this author Theodorus H. Van Der Kwast Toronto, Canada More articles by this author Bas W. Van Rhiyn Amsterdam, Netherlands More articles by this author Joachim W. Thueroff Mainz, Germany More articles by this author Peter C. Black Vancouver, Canada More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.001 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.005 | 0.001 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".