DNMT3B对人脐带间质干细胞成骨分化能力的影响

Abstract

Abstract: 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.

Key words: Osteogenesis, Mesenchymal stem cells, DNA methyltransferase 3B, DNA methylation

CLC Number:

R 681.5+3

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]. 外科研究与新技术, 2012, 1(2): 103-107.

References

[1] Yagi T,Ito D,Okada Y,et al.Modeling familial Alzheimers disease with induced pluripotent stem cells[J].Hum Mol Genet,2011,20(23):4530-4539.
[2] Wernig M,Zhao JP,Pruszak J,et al.Neurons derived from reprogrammed fibroblasts functionally integrate into the fetal brain and improve symptoms of rats with Parkinson’s disease[J].Proc Natl Acad Sci U S A,2008,105(15):5856-5861.
[3] Marchetto MC,Carromeu C,Acab A,et al.A model for neural development and treatment of Rett syndrome using human induced pluripotent stem cells[J].Cell,2010,143(4):527-539.
[4] Urbach A,Bar-Nur O,Daley GQ,et al.Differential modeling of fragile X syndrome by human embryonic stem cells and induced pluripotent stem cells[J].Cell Stem Cell,2010,6(5):407-411.
[5] Hagleitner MM,Lankester A,Maraschio P,et al.Clinical spectrum of immunodeficiency,centromeric instability and facial dysmorphism (ICF syndrome)[J].J Med Genet,2008,45(2):93-99.
[6] Jiang YL,Rigolet M,Bourchis D,et al.DNMT3B mutations and DNA methylation defect define two types of ICF syndrome[J].Hum Mutat,2005,25(1):56-63.
[7] Wu Z,Huang K,Yu J,et al.Dnmt3a regulates both proliferation and differentiation of mouse neural stem cells.J Neurosci Res,2012,90(10):1883-1891.
[8] Watt F and Molloy PL Cytosine methylation prevents binding to DNA of a HeLa cell transcription factor required for optimal expression of the adenovirus major late promoter[J].Genes Dev,1988,2(9):1136-1143.
[9] Lorincz MC,Schubeler D and Groudine M Methylation-mediated proviral silencing is associated with MeCP2 recruitment and localized histone H3 deacetylation[J].Mol Cell Biol,2001,21(23):7913-7922.
[10] Ehrlich M,Sanchez C,Shao C,et al.ICF,an immunodeficiency syndrome:DNA methyltransferase 3B involvement,chromosome anomalies,and gene dysregulation[J].Autoimmunity,2008,41(4):253-271.

DNMT3B regulates osteogenesis of Human Umbilical Mesenchymal Stem Cells

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