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MicroRNA-9 regulates osteoblast differentiation and angiogenesis via the AMPK signaling pathway

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Abstract

MiR-9 has been found to be involved in the repair of spinal cord injury and regulates the proliferation and differentiation of mesenchymal stem cells. However, the role of miR-9 in repair of bone defects has not been well studied. The current study was designed to investigate its role and potential underlying mechanism in regulating osteoblast differentiation and angiogenesis. After treating the murine pre-osteoblast cell line MC3T3-E1 with BMP2, miR-9 expression was obviously down-regulated. Following transfection with miR-9 mimics, its overexpression enhanced the differentiation of MC3T3-E1 cells into osteoblasts as evidence that miR-9 up-regulated the mRNA levels of osteoblast differentiation-related protein, as well as increased differentiation and mineralization of osteoblasts. Further functional analysis has shown that miR-9 overexpression effectively increased human umbilical vein endothelial cell proliferation. Moreover, miR-9 up-regulation promoted cell migration, VEGF, and VE-cadherin concentrations, as well as tube formation in vitro. The mechanistic assay demonstrated that overexpression of miR-9-induced activation of the AMPK signaling pathway. Taken together, our findings suggested that miR-9 overexpression promoted osteoblast differentiation and angiogenesis via the AMPK signaling pathway, representing a novel and potential therapeutic target for the treatment of bone injury-related diseases.

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Abbreviations

BMP2:

Bone morphogenetic protein 2

HUVECs:

Human umbilical vein endothelial cells

VEGF:

Vascular endothelial growth factor

VE-cadherin:

Vascular endothelial cadherin

AMPK:

Adenosine 5′-monophosphate (AMP)-activated protein kinase

iPS:

Induced pluripotent stem cells

MSC:

Mesenchymal stem cell

aMEM:

a-Minimal essential medium

FBS:

Fetal bovine serum

ALP:

Alkaline phosphatase

ARS:

Alizarin red staining

OC:

Osteocalcin

Runx2:

Runt-related transcription factor 2

OSX:

Osterix

p21:

Cyclin-dependent kinase inhibitor 1

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    Authors and Affiliations

    1. Department of Pediatric Orthopedics, Honghui Hospital, Xi’an Jiaotong University Health Science Center, Xi’an, 710054, China

      Jining Qu, Wusheng Miao, Ge Wu & Meifen Zhou

    2. Department of Trauma, Honghui Hospital, Xi’an Jiaotong University Health Science Center, Xi’an, 710054, China

      Daigang Lu

    3. Department of Emergency, Honghui Hospital, Xi’an Jiaotong University Health Science Center, Xi’an, 710054, China

      Hua Guo

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    1. Jining Qu
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    2. Daigang Lu
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    3. Hua Guo
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    Correspondence to Jining Qu or Hua Guo.

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    Jining Qu and Daigang Lu have contributed equally to this work.

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    Qu, J., Lu, D., Guo, H. et al. MicroRNA-9 regulates osteoblast differentiation and angiogenesis via the AMPK signaling pathway. Mol Cell Biochem 411, 23–33 (2016). https://doi.org/10.1007/s11010-015-2565-1

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