Effect of silencing myocardin gene expression on differentiation of mouse bone mesenchymal stem cells into smooth muscle-like cells induced by PDGF-BB / 中华病理学杂志
Chinese Journal of Pathology
; (12): 117-120, 2009.
Article
在 Zh
| WPRIM
| ID: wpr-319775
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>Construction of a small interfering RNA (siRNA) eukaryotic expression vector specific to mouse myocardin gene and study on the role of myocardin-siRNA on differentiation of mouse bone mesenchymal stem cells (MSCs) into smooth muscle-like cells induced by PDGF-BB in vitro.</p><p><b>METHODS</b>Mouse MSCs were isolated from bone marrow and cultured with 50 mg/L PDGF-BB and fetal bovine serum (20%). Specific myocardin-siRNA sequence was cloned into a plasmid pGenesil-1.0 vector, which contained U6 promoter. The recombinant plasmid and control plasmid were transfected into MSCs which had been cultured with PDGF-BB for 6 days beforehand. The expression of myocardin mRNA was detected by RT-PCR 48 hours after the transfection. Immunohistochemistry was used to detect the SM-MHC and to identify the smooth muscle-like cells.</p><p><b>RESULTS</b>The recombinant plasmids carrying myocardin-siRNA sequences were constructed successfully and the myocardin mRNA was reduced 42.86% by pGen-myo-shRNA in comparing with that of the controls (P<0.01); and the expression of SM-MHC protein was down-regulated (P<0.01).</p><p><b>CONCLUSION</b>Subset of mouse MSCs have the potential to differentiate into smooth muscle-like cells, a possible cell source responsible for atherosclerotic plaque formation, and myocardin expression may play an important role during this process.</p>
全文:
1
索引:
WPRIM
主要主题:
Pathology
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Pharmacology
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Physiology
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Plasmids
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Platelet-Derived Growth Factor
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RNA, Messenger
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Bone Marrow Cells
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Nuclear Proteins
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Transfection
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Down-Regulation
研究类型:
Prognostic_studies
限制:
Animals
语言:
Zh
期刊:
Chinese Journal of Pathology
年:
2009
类型:
Article