MiR-9-5p Down-Regulates PiT2, but not PiT1 in Human Embryonic Kidney 293 Cells

Inorganic phosphate (Pi) is an essential component for structure and metabolism. PiT1 ( SLC20A1 ) and PiT2 ( SLC20A2 ) are members of the mammalian type-III inorganic phosphate transporters. SLC20A2 missense variants are associated with primary brain calcification. MicroRNAs (miRNAs) are endogenous...

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Bibliographic Details
Published in:Journal of molecular neuroscience 2017-05, Vol.62 (1), p.28-33
Main Authors: Paiva, D.P., Keasey, M., Oliveira, J. R. M.
Format: Article
Language:English
Subjects:
Affinity
Binding sites
Biomedical and Life Sciences
Biomedicine
Brain
Calcification
Cell Biology
Down-Regulation
Fibrillin
Gene expression
Gene regulation
Genes
HEK293 Cells
Humans
Metabolism
MicroRNAs - genetics
MicroRNAs - metabolism
miRNA
Nervous system
Neurochemistry
Neurogenesis
Neurology
Neurosciences
Phosphate
Phosphates
Pit1 protein
Platelet-derived growth factor
Post-transcription
Proteomics
Ribonucleic acid
RNA
Sodium-Phosphate Cotransporter Proteins, Type III - genetics
Sodium-Phosphate Cotransporter Proteins, Type III - metabolism
Stem cells
Transcription
Western blotting
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Summary:Inorganic phosphate (Pi) is an essential component for structure and metabolism. PiT1 ( SLC20A1 ) and PiT2 ( SLC20A2 ) are members of the mammalian type-III inorganic phosphate transporters. SLC20A2 missense variants are associated with primary brain calcification. MicroRNAs (miRNAs) are endogenous noncoding regulatory RNAs, which play important roles in post-transcriptional gene regulation. MicroRNA-9 (miR-9) acts at different stages of neurogenesis, is deeply rooted in gene networks controlling the regulation of neural progenitor proliferation, and is also linked with cancers outside the nervous system. We evaluated possible interactions between miR-9 and the phosphate transporters (PiT1 and PiT2). SLC20A2, platelet-derived growth factor receptor beta (PDGFRB) and Fibrillin-2 (FBN2) showed binding sites with high affinity for mir-9, in silico . miR-9 mimic was transfected into HEK293 cells and expression confirmed by RT-qPCR. Overexpression of miR-9 in these cells caused a significant reduction in PiT2 and FBN2. PDGFRB appeared to be decreased, but was not significantly down-regulated in our hands. PiT1 showed no significant difference relative to controls. The down-regulation of PiT2 protein by miR-9 was confirmed by western blotting. In conclusion, we showed miR-9 can down-regulate PiT2, in HEK293 cells.
ISSN:0895-8696
1559-1166
DOI:10.1007/s12031-017-0906-0