Artikel
MiR-34a-3p directly targets SMAD4, FRAT1 and BCL2 and alters proliferation and apoptosis of meningioma cells in vitro
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Autoren
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Tamara V. Werner
- Saarland University, Human Genetics, Homburg/Saar, Germany
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Martin Hart
- Saarland University, Human Genetics, Homburg/Saar, Germany
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Ruth Nickels
- Saarland University, Clinical and Experimental Surgery, Homburg/Saar, Germany
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Yoo-Jin Kim
- Saarland University Medical Center, Pathology, Homburg/Saar, Germany
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Michael D. Menger
- Saarland University, Clinical and Experimental Surgery, Homburg/Saar, Germany
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Rainer M. Bohle
- Saarland University Medical Center, Pathology, Homburg/Saar, Germany
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Andreas Keller
- Saarland University, Clinical Bioinformatics, Saarbruecken, Germany
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Nicole Ludwig
- Saarland University, Human Genetics, Homburg/Saar, Germany
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Eckart Meese
- Saarland University, Human Genetics, Homburg/Saar, Germany
| Veröffentlicht: | 14. September 2016 |
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Gliederung
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Introduction: Meningiomas are mostly benign and slow growing tumors of the central nervous system with a small percentage showing a malignant phenotype that is associated with a poor clinical outcome. Micro (mi)RNAs are short, noncoding RNAs that are implicated in the regulation of cellular signaling pathways by targeting and thereby regulating several components of this pathways. Deregulation of miRNAs and their targets lead to dysregulation of signaling pathways, a crucial event in tumor initiation and progression in several cancers.
Objectives: The aim of our study was the identification and validation of targets for miR-34a-3p, a miRNA that is deregulated in higher-grade meningiomas. In addition, we investigated the influence of overexpression or inhibition of miR-34a-3p on meningioma cells in vitro.
Materials and Methods: We performed an in silico analysis for identification of predicted targets for miR-34a-3p and performed luciferase assays with co-expression of miR-34a-3p and reporter gene constructs with the respective 3’ untranslated regions (UTRs). Regulation on protein level was confirmed by Western blotting. The meningioma derived cell line Ben-Men-1 was transfected with synthetic miRNA mimics or inhibitors to investigate the effects of overexpression or inhibition of miR-34a-3p on cell proliferation and apoptosis.
Results: The luciferase assays revealed a negative regulatory effect of miR-34a-3p on the 3’ untranslated regions (UTRs) of SMAD4, FRAT1 and BCL2. Disruption of the miR-34a-3p binding sites in these constructs resulted in loss of responsiveness to miR-34a-3p overexpression. Furthermore, overexpression of miR-34a-3p resulted in a decrease whereas of miR-34a-3p resulted in an increase of SMAD4, FRAT1 and BCL2 protein levels in meningioma cells as confirmed by Western blotting. Deregulation of miR-34a-3p altered cell proliferation and apoptosis of the meningioma cell line Ben-Men-1.
Conclusion: These findings support the hypothesis that loss of translational repression of SMAD4, FRAT1 and BCL2 due to downregulation of miR-34a-3p may contribute to processes that are crucial for tumor initiation and progression, e.g. epithelial-to-mesenchymal transition (EMT), proliferation, invasion, migration and apoptosis.
