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Updated: Jun 3, 2026
High-throughput Screening for Chemical Modulators of Post-transcriptionally Regulated Genes
Published on: March 3, 2015
Identification and validation of Notch pathway activating compounds through a novel high-throughput screening method
Scott N Pinchot1, Renata Jaskula-Sztul, Li Ning
1Endocrine Surgery Research Laboratory, Department of Surgery and the University of Wisconsin Carbone Cancer Center, Madison, Wisconsin 53792, USA.
Background:
Carcinoids are neuroendocrine (NE) tumors with limited treatment options. Notch activation has been shown to suppress growth and hormone production in carcinoid cells.
Methods:
The purpose of this study was to provide a process for identifying Notch activating compounds via high-throughput screening (HTS) and to validate the effects of the strongest hit from the 7264 compounds analyzed: resveratrol (RESV).
Results:
Treatment of carcinoid cells with RESV resulted in up-regulation of the Notch signaling pathway as measured by suppression of its downstream target achaete-scute complex-like 1. Luciferase reporter assays incorporating the centromere-binding factor 1 binding site also confirmed the functional activity of RESV-induced Notch. Because activation of the Notch pathway has been shown to suppress carcinoid proliferation, RESV treatment of carcinoid cells led to a dose-dependent inhibition of cellular growth. Immunoblotting revealed phosphorylation of cdc2 (Tyr15) and up-regulation of p21Cip1/Waf, markers of cell cycle arrest, with RESV treatment. Flow cytometry confirmed the mechanism of RESV-induced growth inhibition is S phase cell cycle arrest. Furthermore, because Notch has been shown to inhibit bioactive hormone production from NE tumors, RESV also suppressed expression of the NE peptides/hormones chromogranin A and serotonin. RNA interference assays demonstrated that the hormone suppressing capacity of RESV was due to up-regulation of the Notch2 isoform.
Conclusions:
HTS can be used to identify novel Notch activating compounds, which may have the potential to suppress carcinoid tumor growth and the associated endocrinopathies. Cancer 2011. © 2010 American Cancer Society.
Insights
High-throughput screening identified resveratrol as a compound that activates Notch signaling. This activation suppresses carcinoid tumor growth and hormone production, offering a potential new treatment strategy.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Carcinoid tumors are neuroendocrine (NE) neoplasms with limited therapeutic options.
- Notch pathway activation demonstrates potential for suppressing carcinoid cell growth and hormone secretion.
Purpose of the Study:
- To establish a high-throughput screening (HTS) process for identifying Notch-activating compounds.
- To validate the efficacy of resveratrol (RESV), the strongest compound identified, in carcinoid cells.
Main Methods:
- Utilized HTS to analyze 7264 compounds for Notch pathway activation.
- Employed cell-based assays, including luciferase reporter assays and immunoblotting, to assess pathway modulation and cellular effects.
- Conducted flow cytometry and RNA interference assays to elucidate mechanisms of action.
Main Results:
- Resveratrol (RESV) treatment upregulated Notch signaling, evidenced by suppressed expression of its target gene, achaete-scute complex-like 1.
- RESV induced a dose-dependent inhibition of carcinoid cell proliferation via S phase cell cycle arrest, confirmed by markers like p21Cip1/Waf.
- RESV suppressed the production of neuroendocrine peptides, including chromogranin A and serotonin, mediated by Notch2 isoform upregulation.
Conclusions:
- High-throughput screening is effective for discovering novel Notch-activating compounds for carcinoid therapy.
- Resveratrol demonstrates potential as a therapeutic agent for carcinoid tumors by inhibiting growth and hormone production.
- Targeting the Notch pathway offers a promising strategy to manage carcinoid tumors and associated endocrinopathies.
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