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Updated: Apr 14, 2026
High-throughput Screening for Chemical Modulators of Post-transcriptionally Regulated Genes
Published on: March 3, 2015
Identification of novel pathways and molecules able to down-regulate PHOX2B gene expression by in vitro drug
Eleonora Di Zanni1, Diego Fornasari2, Roberto Ravazzolo3
1U.O.C. Genetica Medica, Istituto Giannina Gaslini, Italy.
Abstract:
PHOX2B is a transcription factor involved in the regulation of neurogenesis and in the correct differentiation of the autonomic nervous system. The pathogenetic role of PHOX2B in neuroblastoma (NB) is supported by mutations in familial, sporadic and syndromic cases of NB and overexpression of PHOX2B and its target ALK in tumor samples and NB cell lines. Starting from these observations, we have performed in vitro drug screening approaches targeting PHOX2B overexpression as a potential pharmacological means in NB. In particular, in order to identify molecules able to decrease PHOX2B expression, we have evaluated the effects of 70 compounds in IMR-32 cell line stably expressing the luciferase gene under the control of the PHOX2B promoter. Curcumin, SAHA and trichostatin A showed to down-regulate the PHOX2B promoter activity which resulted in a decrease of both protein and mRNA expressions. In addition, we have observed that curcumin acts by interfering with PBX-1/MEIS-1, NF-κB and AP-1 complexes, in this work demonstrated for the first time to regulate the transcription of the PHOX2B gene. Finally, combined drug treatments showed successful effects in down-regulating the expression of both PHOX2B and its target ALK genes, thus supporting the notion of the effectiveness of molecule combination in tumor therapy.
Insights
This study screened compounds to target PHOX2B overexpression in neuroblastoma (NB). Curcumin, SAHA, and trichostatin A reduced PHOX2B expression, with curcumin affecting key transcription factors, showing potential for combination therapy.
Area of Science:
- Molecular Biology
- Genetics
- Oncology
Background:
- PHOX2B is a crucial transcription factor for autonomic nervous system development.
- PHOX2B mutations and overexpression are implicated in neuroblastoma (NB) pathogenesis.
- Overexpression of PHOX2B and its target ALK is observed in NB tumors.
Purpose of the Study:
- To identify compounds that down-regulate PHOX2B expression as a therapeutic strategy for NB.
- To investigate the molecular mechanisms by which these compounds affect PHOX2B transcription.
- To evaluate the efficacy of combined drug treatments targeting PHOX2B and ALK.
Main Methods:
- In vitro drug screening of 70 compounds using a PHOX2B promoter-luciferase reporter assay in IMR-32 NB cells.
- Analysis of PHOX2B protein and mRNA levels following compound treatment.
- Investigation of curcumin's effect on transcription factor complexes (PBX-1/MEIS-1, NF-κB, AP-1).
- Assessment of combined drug treatment effects on PHOX2B and ALK expression.
Main Results:
- Curcumin, SAHA, and trichostatin A significantly down-regulated PHOX2B promoter activity, protein, and mRNA expression.
- Curcumin was found to interfere with PBX-1/MEIS-1, NF-κB, and AP-1 transcription factor complexes regulating PHOX2B.
- Combined drug treatments effectively reduced the expression of both PHOX2B and its target ALK.
Conclusions:
- PHOX2B is a viable therapeutic target in neuroblastoma.
- Curcumin, SAHA, and trichostatin A show promise in down-regulating PHOX2B expression.
- Combination therapy targeting PHOX2B and ALK offers a potential strategy for NB treatment.
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