Dataset Information

Long noncoding RNA LINC00707 sponges miR-370-3p to promote osteogenesis of human bone marrow-derived mesenchymal stem cells through upregulating WNT2B.

ABSTRACT:

Background

Human bone marrow-derived mesenchymal stem cells (HBMSCs) are characterized by multiple differentiation potential and potent self-renewal ability, yet much remains to be elucidated on what determines these properties. Long-chain noncoding RNAs (lncRNAs) have been suggested to be involved in multiple biological processes under physiological and pathological conditions, including osteogenic differentiation.

Methods

Alkaline phosphatase (ALP) activity assay, ALP staining, and Alizarin Red Staining were used for osteogenic potential detection. Western blot and qRT-PCR were used to examine the expression of LINC00707 and miR-370-3p. RNA-binding protein immunoprecipitation was used to detect the interaction between LINC00707 and RNA-induced silencing complex. Luciferase reporter assay was used to confirm the binding sites of miR-370-3p to LINC00707 and WNT2B.

Results

We demonstrated that LINC00707 expression was gradually increased in HBMSCs during consecutive osteogenic induction, and it could further positively regulate the osteogenic differentiation both in vitro and in vivo, whereas LINC00707 inhibition led to suppressed osteogenic differentiation. Thereafter, we inferred a predicted interaction between LINC00707 and miR-370-3p and then confirmed the direct binding sites of miR-370-3p on LINC00707. While miR-370-3p upregulation led to decreased osteogenic differentiation, LINC00707 overexpression could reverse this suppression, indicating that LINC00707 acts as a competing endogenous RNA (ceRNA) for miR-370-3p. Moreover, LINC00707 could act as a ceRNA to upregulate WNT2B via miR-370-3p inhibition.

Conclusions

In conclusion, our study provides a novel lncRNA-miRNA regulatory network and a promising target to modulate the osteogenic differentiation of HBMSCs.

SUBMITTER: Jia B

PROVIDER: S-EPMC6387535 | biostudies-literature | 2019 Feb

REPOSITORIES: biostudies-literature

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Publications

Long noncoding RNA LINC00707 sponges miR-370-3p to promote osteogenesis of human bone marrow-derived mesenchymal stem cells through upregulating WNT2B.

Jia Bo B Wang Zhiping Z Sun Xiang X Chen Jun J Zhao Jianjiang J Qiu Xiaoling X

Stem cell research & therapy 20190222 1

<h4>Background</h4>Human bone marrow-derived mesenchymal stem cells (HBMSCs) are characterized by multiple differentiation potential and potent self-renewal ability, yet much remains to be elucidated on what determines these properties. Long-chain noncoding RNAs (lncRNAs) have been suggested to be involved in multiple biological processes under physiological and pathological conditions, including osteogenic differentiation.<h4>Methods</h4>Alkaline phosphatase (ALP) activity assay, ALP staining, ...[more]

PMID: 30795799

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Project description:BackgroundTissue-engineered bone grafts (TEBGs) that undergo vascularization and neurotization evolve into functioning bone tissue. Previously, we verified that implanting sensory nerve tracts into TEBGs promoted osteogenesis. However, the precise mechanisms and interaction between seed cells were not explored. In this study, we hypothesized that neurotization may influence the osteogenesis of TEBGs through vascularization.MethodsWe cultured rat Schwann cells (SCs), aortic endothelial cells (AECs), and bone marrow-derived mesenchymal stem cells (BM-MSCs) and then obtained BM-MSC-derived induced endothelial cells (IECs) and induced osteoblasts (IOBs). IECs and AECs were cultured in an SC-conditioned medium (SC-CM) to assess proliferation, migration, capillary-like tube formation, and angiogenesis, and the vascular endothelial growth factor (VEGF) levels in the supernatants were detected. We established an indirect coculture model to detect the expression of nestin and VEGF receptors in IECs and tissue inhibitor of metalloproteinase (TIMP)-2 in SCs. Then, SCs, IECs, and IOBs were labeled and loaded into a β-tricalcium phosphate scaffold to induce prevascularization, and the scaffold was implanted into a 6-mm-long defect of rat femurs. Three groups were set up according to the loaded cells: I, SCs, and IECs (coculture for 3 days) plus IOBs; II, IECs (culture for 3 days) plus IOBs; III, IOBs. Nestin and TIMP-2 expression and osteogenesis of TEBGs were evaluated at 12 weeks post-implantation through histological and radiological assessments.ResultsWe found that SC-CM promoted IEC proliferation, migration, capillary-like tube formation, and angiogenesis, but no similar effects were observed for AECs. IECs expressed nestin extensively, while AECs barely expressed nestin, and SC-CM promoted the VEGF secretion of IECs. In the coculture model, SCs promoted nestin and VEGF receptor expression in IECs, and IECs inhibited TIMP-2 expression in SCs. The promotion of prevascularized TEBGs by SCs and IECs in group I augmented new bone formation at 6 and 12 weeks. Nestin expression was higher in group I than in the other groups, while TIMP-2 expression was lower at 12 weeks.ConclusionsThis study demonstrated that SCs can promote TEBG osteogenesis via IECs and further revealed the related specific characteristics of IECs, providing preliminary cytological evidence for neurotization of TEBGs.

Dataset Information

Long noncoding RNA LINC00707 sponges miR-370-3p to promote osteogenesis of human bone marrow-derived mesenchymal stem cells through upregulating WNT2B.

Background

Methods

Results

Conclusions

Publications

Long noncoding RNA LINC00707 sponges miR-370-3p to promote osteogenesis of human bone marrow-derived mesenchymal stem cells through upregulating WNT2B.

Similar Datasets

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