Abstract
Micro (mi)RNAs are short, noncoding RNAs and deregulation of miRNAs and their targets are implicated in tumor generation and progression in many cancers. Meningiomas are mostly benign, slow growing tumors of the central nervous system with a small percentage showing a malignant phenotype.Following in silico prediction of potential targets of miR-34a-3p, SMAD4, FRAT1, and BCL2 have been confirmed as targets by dual luciferase assays with co-expression of miR-34a-3p and reporter gene constructs containing the respective 3'UTRs. Disruption of the miR-34a-3p binding sites in the 3'UTRs resulted in loss of responsiveness to miR-34a-3p overexpression. In meningioma cells, overexpression of miR-34a-3p resulted in decreased protein levels of SMAD4, FRAT1 and BCL2, while inhibition of miR-34a-3p led to increased levels of these proteins as confirmed by Western blotting. Furthermore, deregulation of miR-34a-3p altered cell proliferation and apoptosis of meningioma cells in vitro.We show that SMAD4, FRAT1 and BCL2 are direct targets of miR-34a-3p and that deregulation of miR-34a-3p alters proliferation and apoptosis of meningioma cells in vitro. As part of their respective signaling pathways, which are known to play a role in meningioma genesis and progression, deregulation of SMAD4, FRAT1 and BCL2 might contribute to the aberrant activation of these signaling pathways leading to increased proliferation and inhibition of apoptosis in meningiomas.
Highlights
Protein levels were semiquantified using the Immuno-Reactive-Score (IRS). (A, B) Representative images for frequently rearranged in advanced T-cell lymphomas 1 (FRAT1) staining for samples with an IRS of 4 (A) and 12 (B). (C, D) Representative images for SMAD family member 4 (SMAD4) staining with an IRS of 4 (C) and 8 (D)
The stop codon is highlighted in red, the potential binding site for miR-34a-3p is highlighted in green and the complete fragment cloned into pMIR-RNL-TK is underlined
Homo sapiens frequently rearranged in advanced T-cell lymphomas 1 (FRAT1), mRNA
Summary
Protein levels were semiquantified using the Immuno-Reactive-Score (IRS). (A, B) Representative images for FRAT1 staining for samples with an IRS of 4 (A) and 12 (B). (C, D) Representative images for SMAD4 staining with an IRS of 4 (C) and 8 (D).
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