Dataset Information

MicroRNA regulation of neuron-like differentiation of adipose tissue-derived stem cells.

ABSTRACT: We have reported previously that adipose tissue-derived stem cells (ADSC) could be induced by isobutylmethylxanthine (IBMX) to differentiate into neuron-like cells and such differentiation was mediated by insulin-like growth factor I (IGF-I) signaling. In the present study we show that both IBMX and IGF-I upregulated neural markers beta-III-tubulin and paired-like homeobox transcription factor (Pitx3) in ADSC. Because Pitx3 is important for the maturation and function of dopaminergic neurons and has been shown to be regulated by microRNA miR-133b, we also examined the effects of IBMX and IGF-I on miR-133b expression. The results show that both IBMX and IGF-I upregulated miR-133b expression. When miR-133b was overexpressed in ADSC through transfection of a synthetic microRNA mimic, it caused a downregulation of beta-III-tubulin as evidenced by immunofluorescence staining and western blot analysis. Overexpression of miR-133b also downregulated Pitx3 and IGF-1 receptor (IGF-IR), and all such downregulation occurred at the protein but not the RNA level. The apparent posttranscriptional regulation was subsequently found to be exerted through a potential miR-133b target in the 3'-untranslated region of IGF-IR, as evidenced by luciferase assay. Thus, IGF-I signaling and miR-133b co-regulate potential neural differentiation of ADSC through a feedback mechanism, in which IGF-I upregulates miR-133b while miR-133b in turn downregulates IGF-IR.

SUBMITTER: Ning H

PROVIDER: S-EPMC2787638 | biostudies-literature | 2009 Dec

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

MicroRNA regulation of neuron-like differentiation of adipose tissue-derived stem cells.

Ning Hongxiu H Huang Yun-Ching YC Banie Lia L Hung Steven S Lin Guiting G Li Long-Cheng LC Lue Tom F TF Lin Ching-Shwun CS

Differentiation; research in biological diversity 20090819 5

We have reported previously that adipose tissue-derived stem cells (ADSC) could be induced by isobutylmethylxanthine (IBMX) to differentiate into neuron-like cells and such differentiation was mediated by insulin-like growth factor I (IGF-I) signaling. In the present study we show that both IBMX and IGF-I upregulated neural markers beta-III-tubulin and paired-like homeobox transcription factor (Pitx3) in ADSC. Because Pitx3 is important for the maturation and function of dopaminergic neurons and ...[more]

PMID: 19695767

Similar Datasets

Project description:BACKGROUND:MicroRNAs (miRNAs) play a role in regulating osteogenic differentiation (OD) of mesenchymal stem cells by inhibiting mRNAs translation under cyclic strain. miR-503-3p was downregulated in OD of human adipose-derived stem cells (hASCs) in vivo under cyclic strain in our previous study, while it might target the Wnt/?-catenin (W-?) pathway. In this study, we explored miR-503-3p's role in OD of hASCs under cyclic strain. METHODS:OD of hASCs was induced by cyclic strain. Bioinformatic and dual luciferase analyses were used to confirm the relationship between Wnt2/Wnt7b and miR-503-3p. Immunofluorescence was used to detect the effect of miR-503-3p on Wnt2/Wnt7b and ?-catenin in hASCs transfected with miR-503-3p mimic and inhibitor. Mimic, inhibitor, and small interfering RNA (siRNA) transfected in hASCs to against Wnt2 and Wnt7b. Quantitative real-time PCR (RT-PCR) and western blot were used to examine the OD and W-? pathway at the mRNA and protein levels, respectively. Immunofluorescence was performed to locate ?-catenin. ALP activity and calcium were detected by colorimetric assay. RESULTS:Results of immunophenotypes by flow cytometry and multi-lineage potential confirmed that the cultured cells were hASCs. Results of luciferase reporter assay indicated that miR-503-3p could regulate the expression levels of Wnt2 and Wnt7b by targeting their respective 3'-untranslated region (UTR). Under cyclic strain, gain- or loss-function of miR-503-3p studies by mimic and inhibitor revealed that decreasing expression of miR-503-3p could significantly bring about promotion of OD of hASCs, whereas increased expression of miR-503-3p inhibited OD. Furthermore, miR-503-3p high-expression reduced the activity of the W-? pathway, as indicated by lowering expression of Wnt2 and Wnt7b, inactive ?-catenin in miR-503-3p-treated hASCs. By contrast, miR-503-3p inhibition activated the W-? pathway. CONCLUSIONS:Collectively, our findings indicate that miR-503-3p is a negative factor in regulating W-? pathway by Wnt2 and Wnt7b, which inhibit the OD of hASCs under cyclic strain.

Dataset Information

MicroRNA regulation of neuron-like differentiation of adipose tissue-derived stem cells.

Publications

MicroRNA regulation of neuron-like differentiation of adipose tissue-derived stem cells.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets