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
in Chinese
| WPRIM
| ID: wpr-319775
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>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pathology
/
Pharmacology
/
Physiology
/
Plasmids
/
Platelet-Derived Growth Factor
/
RNA, Messenger
/
Bone Marrow Cells
/
Nuclear Proteins
/
Transfection
/
Down-Regulation
Type of study:
Prognostic study
Limits:
Animals
Language:
Chinese
Journal:
Chinese Journal of Pathology
Year:
2009
Type:
Article
Similar
MEDLINE
...
LILACS
LIS