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The Effect of EPO Gene Overexpression on Proliferation and Migration of Mouse Bone Marrow-Derived Mesenchymal Stem Cells

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Abstract

The aim of this study is to investigate the effect of erythropoietin (EPO) gene overexpression on proliferation and migration of mouse bone marrow-derived mesenchymal stem cells (MSCs), and to determine the underlying signaling pathway. Mouse MSCs were cultured in vitro and EPO gene was transfected into the 6th generation of MSCs via lentivirus vector. The transfected cells were identified by flow cytometry and the EPO levels in supernatant were measured with ELISA. In addition, cell proliferation was assessed by CCK-8 assay and cell migration was evaluated by Transwell assay. The activation of Akt, ERK1/2, and p38MAPK signaling was detected by western blotting. The lentivirus vector containing EPO was successfully constructed and transfected into MSCs. No remarkable change was found in the cell surface markers after transfection while a significant increase of EPO level in supernatant was noticed in transfected MSCs compared to controls (P < 0.01). In addition, transfected MSCs showed a significantly enhanced proliferation (P < 0.01) as well as a notable increase in migration (P < 0.01) compared to controls. Furthermore, we also found that EPO modification enhanced the phosphorylation of PI3K/Akt and ERK signaling pathway, and suppressed the phosphorylation of p38MAPK without affecting the levels of total Akt, ERK1/2, and p38MAPK in MSCs. After transfection, MSCs secreted more EPO which enhanced the capability of proliferation and migration. Moreover, our results suggested that the enhanced proliferation and migration might be associated with activation of PI3K/Akt and ERK or inhibition of P38MAPK signaling pathway.

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Authors and Affiliations

  1. Department of Cardiology, Jinshan Hospital Affiliated to Fudan University, Shanghai, 200433, China

    Haihong Lin & Hui Gong

  2. Department of Cardiology, Huashan Hospital Affiliated to Fudan University, Shanghai, 200433, China

    Xinping Luo, Bo Jin & Haiming Shi

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  1. Haihong Lin
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  2. Xinping Luo
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  3. Bo Jin
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  4. Haiming Shi
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  5. Hui Gong
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Corresponding author

Correspondence to Hui Gong.

Additional information

Dr. Haihong Lin and Dr. Xinping Luo have contributed equally to this study.

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Lin, H., Luo, X., Jin, B. et al. The Effect of EPO Gene Overexpression on Proliferation and Migration of Mouse Bone Marrow-Derived Mesenchymal Stem Cells. Cell Biochem Biophys 71, 1365–1372 (2015). https://doi.org/10.1007/s12013-014-0358-x

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