Enhanced mesenchymal stem cell survival induced by GATA-4 overexpression is partially mediated by regulation of the miR-15 family

Bin Yu, Min Gong, Zhisong He, Yi Gang Wang, Ronald W. Millard, Muhammad Ashraf, Meifeng Xu

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

We reported previously that pre-programming mesenchymal stem cells with the GATA-4 gene increases significantly cell survival in an ischemic environment. In this study, we tested whether regulation of microRNAs and their target proteins was associated with the cytoprotective effects of GATA-4. Methods and results Mesenchymal stem cells were harvested from adult rat bone marrow and transduced with GATA-4 (MSCGATA-4) using the murine stem cell virus retroviral expression system. Cells transfected with empty vector (MSC Null) were used as controls. Quantitative real-time PCR data showed that the expression levels of miR-15 family members (miR-15b, miR-16, and miR-195) were significantly down-regulated in MSCGATA-4. The protein expression of Bcl-w (Bcl-2-like-2), an anti-apoptotic Bcl-2 family protein, was increased in MSCGATA-4. Hypoxic culture (low glucose and low oxygen) induced the release of lactate dehydrogenase from mesenchymal stem cells and reduced cell survival. Compared to MSCNull, MSCGATA-4 showed less lactate dehydrogenase release and greater cell survival following 72 h hypoxia exposure. The mitochondrial membrane potential, detected with the dye JC-1, was well maintained, and mitochondrial membrane permeability, expressed as caspase 3 and 7 activities in response to the ischemic environment was lower in MSCGATA-4. Moreover, transfection with miR-195 significantly down-regulated Bcl-w expression in mesenchymal stem cells through a binding site in the 3′-UTR of the Bcl-w mRNA and reduced mesenchymal stem cell resistance to ischemic injury. Conclusions The overexpression of GATA-4 in mesenchymal stem cells down-regulates miR-15 family members, causing increased resistance to ischemia through the up-regulation of anti-apoptotic proteins in the Bcl-2 family.

Original languageEnglish (US)
Pages (from-to)2724-2735
Number of pages12
JournalInternational Journal of Biochemistry and Cell Biology
Volume45
Issue number12
DOIs
StatePublished - Oct 28 2013

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Stem cells
Mesenchymal Stromal Cells
Cell Survival
Bone Marrow
L-Lactate Dehydrogenase
Cells
Caspase 7
Apoptosis Regulatory Proteins
Proteins
Membranes
Mitochondrial Membrane Potential
Mitochondrial Membranes
3' Untranslated Regions
MicroRNAs
Caspase 3
Transfection
Real-Time Polymerase Chain Reaction
Viruses
Permeability
Coloring Agents

Keywords

  • Bcl-w
  • Cytoprotection
  • GATA-4
  • Mesenchymal stem cells
  • miR-15 family

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

Enhanced mesenchymal stem cell survival induced by GATA-4 overexpression is partially mediated by regulation of the miR-15 family. / Yu, Bin; Gong, Min; He, Zhisong; Wang, Yi Gang; Millard, Ronald W.; Ashraf, Muhammad; Xu, Meifeng.

In: International Journal of Biochemistry and Cell Biology, Vol. 45, No. 12, 28.10.2013, p. 2724-2735.

Research output: Contribution to journalArticle

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abstract = "We reported previously that pre-programming mesenchymal stem cells with the GATA-4 gene increases significantly cell survival in an ischemic environment. In this study, we tested whether regulation of microRNAs and their target proteins was associated with the cytoprotective effects of GATA-4. Methods and results Mesenchymal stem cells were harvested from adult rat bone marrow and transduced with GATA-4 (MSCGATA-4) using the murine stem cell virus retroviral expression system. Cells transfected with empty vector (MSC Null) were used as controls. Quantitative real-time PCR data showed that the expression levels of miR-15 family members (miR-15b, miR-16, and miR-195) were significantly down-regulated in MSCGATA-4. The protein expression of Bcl-w (Bcl-2-like-2), an anti-apoptotic Bcl-2 family protein, was increased in MSCGATA-4. Hypoxic culture (low glucose and low oxygen) induced the release of lactate dehydrogenase from mesenchymal stem cells and reduced cell survival. Compared to MSCNull, MSCGATA-4 showed less lactate dehydrogenase release and greater cell survival following 72 h hypoxia exposure. The mitochondrial membrane potential, detected with the dye JC-1, was well maintained, and mitochondrial membrane permeability, expressed as caspase 3 and 7 activities in response to the ischemic environment was lower in MSCGATA-4. Moreover, transfection with miR-195 significantly down-regulated Bcl-w expression in mesenchymal stem cells through a binding site in the 3′-UTR of the Bcl-w mRNA and reduced mesenchymal stem cell resistance to ischemic injury. Conclusions The overexpression of GATA-4 in mesenchymal stem cells down-regulates miR-15 family members, causing increased resistance to ischemia through the up-regulation of anti-apoptotic proteins in the Bcl-2 family.",
keywords = "Bcl-w, Cytoprotection, GATA-4, Mesenchymal stem cells, miR-15 family",
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T1 - Enhanced mesenchymal stem cell survival induced by GATA-4 overexpression is partially mediated by regulation of the miR-15 family

AU - Yu, Bin

AU - Gong, Min

AU - He, Zhisong

AU - Wang, Yi Gang

AU - Millard, Ronald W.

AU - Ashraf, Muhammad

AU - Xu, Meifeng

PY - 2013/10/28

Y1 - 2013/10/28

N2 - We reported previously that pre-programming mesenchymal stem cells with the GATA-4 gene increases significantly cell survival in an ischemic environment. In this study, we tested whether regulation of microRNAs and their target proteins was associated with the cytoprotective effects of GATA-4. Methods and results Mesenchymal stem cells were harvested from adult rat bone marrow and transduced with GATA-4 (MSCGATA-4) using the murine stem cell virus retroviral expression system. Cells transfected with empty vector (MSC Null) were used as controls. Quantitative real-time PCR data showed that the expression levels of miR-15 family members (miR-15b, miR-16, and miR-195) were significantly down-regulated in MSCGATA-4. The protein expression of Bcl-w (Bcl-2-like-2), an anti-apoptotic Bcl-2 family protein, was increased in MSCGATA-4. Hypoxic culture (low glucose and low oxygen) induced the release of lactate dehydrogenase from mesenchymal stem cells and reduced cell survival. Compared to MSCNull, MSCGATA-4 showed less lactate dehydrogenase release and greater cell survival following 72 h hypoxia exposure. The mitochondrial membrane potential, detected with the dye JC-1, was well maintained, and mitochondrial membrane permeability, expressed as caspase 3 and 7 activities in response to the ischemic environment was lower in MSCGATA-4. Moreover, transfection with miR-195 significantly down-regulated Bcl-w expression in mesenchymal stem cells through a binding site in the 3′-UTR of the Bcl-w mRNA and reduced mesenchymal stem cell resistance to ischemic injury. Conclusions The overexpression of GATA-4 in mesenchymal stem cells down-regulates miR-15 family members, causing increased resistance to ischemia through the up-regulation of anti-apoptotic proteins in the Bcl-2 family.

AB - We reported previously that pre-programming mesenchymal stem cells with the GATA-4 gene increases significantly cell survival in an ischemic environment. In this study, we tested whether regulation of microRNAs and their target proteins was associated with the cytoprotective effects of GATA-4. Methods and results Mesenchymal stem cells were harvested from adult rat bone marrow and transduced with GATA-4 (MSCGATA-4) using the murine stem cell virus retroviral expression system. Cells transfected with empty vector (MSC Null) were used as controls. Quantitative real-time PCR data showed that the expression levels of miR-15 family members (miR-15b, miR-16, and miR-195) were significantly down-regulated in MSCGATA-4. The protein expression of Bcl-w (Bcl-2-like-2), an anti-apoptotic Bcl-2 family protein, was increased in MSCGATA-4. Hypoxic culture (low glucose and low oxygen) induced the release of lactate dehydrogenase from mesenchymal stem cells and reduced cell survival. Compared to MSCNull, MSCGATA-4 showed less lactate dehydrogenase release and greater cell survival following 72 h hypoxia exposure. The mitochondrial membrane potential, detected with the dye JC-1, was well maintained, and mitochondrial membrane permeability, expressed as caspase 3 and 7 activities in response to the ischemic environment was lower in MSCGATA-4. Moreover, transfection with miR-195 significantly down-regulated Bcl-w expression in mesenchymal stem cells through a binding site in the 3′-UTR of the Bcl-w mRNA and reduced mesenchymal stem cell resistance to ischemic injury. Conclusions The overexpression of GATA-4 in mesenchymal stem cells down-regulates miR-15 family members, causing increased resistance to ischemia through the up-regulation of anti-apoptotic proteins in the Bcl-2 family.

KW - Bcl-w

KW - Cytoprotection

KW - GATA-4

KW - Mesenchymal stem cells

KW - miR-15 family

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