Objective To elucidate the roles of DNMT3B in the osteogenesis of hUMSCs.Methods Knockdown DNMT3B in hUMSCs was gained via RNA interference technology.After confirming the decrease of DNMT3B in mutant hUMSCs by immunostaining and qPCR, osteogenesis differentiation was carried out.To identify the phenotype of osteogenesis in both bone formation ability and function of bone,immunostaining,qPCR and functional test was preformed,compared to widetype hUMSCs.Results The mutant hUMSCs showed lower DNMT3B expression level by qPCR.Compared to widetype cells,the DNMT3B expression level were 12.67±0.45% and 35.60±0.76%in two mutant hUMSCs (P<0.05).Furthermore,the expression level of CD73 and CD90 that were two mesenchymal specific markers were 15.83±0.44% and 16.60±0.37% of widetype hUMSCs (P<0.05).Immunostaining showed Osteocalcin positive cell was significant less in DNMT3B mutant cells.qPCR demonstrated that RUNX2,IBSP and ALPL mRNA level were decreased in mutant cells (P<0.01).Moreover,alizarin red staining performed that functional impairment lacking of DNMT3B in hUMSCs.Conclusion The osteogenesis differentiation of hUMSCs is impaired in the absence of DNMT3B.
Wu Zhou-Rui
,
Hu Xiao
,
Xu Wei
,
Sun Xiao-Qing
,
Guan Xiao-Fei
,
Cheng Li-Ming
. DNMT3B regulates osteogenesis of Human Umbilical Mesenchymal Stem Cells[J]. Surgical Research and New Technique, 2012
, 1(2)
: 103
-107
.
DOI: 10.3969/j.issn.2095-378X.2012.02.003
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