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Published on: March 30, 2019
Recognizing genes differentially regulated in vitro by the multiple endocrine neoplasia type 1 (MEN1) gene, using RNA
Peter Stålberg1, Mårten Santesson, Sara Ekeblad
1Department of Surgical Sciences, University Hospital, Uppsala, Sweden.
Background:
Data on downstream effects of MEN1 gene inactivation is scarce. In an effort to identify genes regulated by MEN1, we designed a silencing experiment in a human endocrine pancreatic tumor cell line (BON1).
Methods:
By using RNA interference, MEN1 mRNA expression was knocked-down by >85%. Gene expression was assessed by oligonucleotide microarrays and compared to expression in nonsilenced controls. We also investigated if genes were differentially expressed in 6 malignant endocrine pancreatic tumors (EPTs) with homozygous MEN1 inactivation compared to 2 without MEN1 gene alterations.
Results:
Using a cut-off of > or =2 times, 66 genes were found to be upregulated, and 22 were downregulated in the MEN1-silenced clones. We corroborated the microarray findings by performing quantitative-PCR on the RNA from the silencing experiments for 7 of the 88 differentially regulated genes. Genes involved in endocrine cell fate determination, as well as genes known to be involved in NFkappaB, Notch, and Wnt signaling pathways, were among genes verified as differentially regulated in vitro.
Conclusions:
The demonstration of pathways affected by silencing of MEN1 in vitro provides novel insight into neoplastic processes of potential importance in vivo, which warrants further study.
Insights
Silencing the MEN1 gene in pancreatic tumor cells revealed 88 differentially expressed genes, including those in key cell signaling pathways. This offers new insights into endocrine tumor development.
Area of Science:
- Endocrinology
- Molecular Biology
- Oncology
Background:
- Downstream effects of MEN1 gene inactivation are not well understood.
- The MEN1 gene is implicated in endocrine tumors.
- Limited data exists on genes regulated by MEN1.
Purpose of the Study:
- To identify genes regulated by MEN1.
- To investigate the downstream effects of MEN1 inactivation.
- To explore signaling pathways affected by MEN1.
Main Methods:
- Utilized RNA interference to knock down MEN1 mRNA expression in a human endocrine pancreatic tumor cell line (BON1).
- Assessed gene expression using oligonucleotide microarrays and quantitative PCR.
- Compared gene expression in silenced cells versus controls and in tumors with and without MEN1 alterations.
Main Results:
- Successfully knocked down MEN1 mRNA expression by over 85%.
- Identified 66 upregulated and 22 downregulated genes in MEN1-silenced cells.
- Confirmed differential regulation of genes involved in endocrine cell fate and NFkappaB, Notch, and Wnt signaling pathways.
Conclusions:
- MEN1 silencing affects key cellular pathways, providing novel insights into neoplastic processes.
- The identified pathways offer potential targets for understanding and treating endocrine tumors.
- Further in vivo studies are warranted to validate these in vitro findings.
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