Wnt/β-catenin pathway might underlie the MET in trans-differentiation from MSC to MSC-derived neuron

【作者】 景莹； 张嘉诚； 李素停； 赵纪华； 王晋； 韩雪飞； 邢莹；

【机构】 郑州大学干细胞研究中心； 郑州大学生理学与神经生物学教研室； 新乡医学院；

【摘要】 Mesenchymal-epithelial transition(MET)occurs commonly in both physiological and pathological context.Objective:To observe MET-associated alteration during the trans-differentiation from MSCs to neuron-like cells,and to explore the possible molecular mechanism.Methods:Bone marrow MSCs were isolated from rat femur and purified in continuous cell culture.After induced differentiation to neuron-like cells by the combination of butylated hydroxyanisole(BHA)and dimethyl sulfoxide(DMSO),cells were tested by comparative polymerase chain reaction(PCR)for the relative expression of MET biomarkers and transcription factors,and for cell cycle by flow cytometry.Meanwhile,target genes of Wnt/β-catenin pathway were also analyzed by comparative PCR to determine the possible involvement.Results:in MSC-induced neuron-like cells,MET-associated transcription factors such as Snail,Slug,ZEB1,ZEB2,and Twist were significantly attenuated in expression level.Mesenchymal marker vimentin was increased.Membrane protein E-cad was slightly down-regulated,while N-cad level was marginally elevated.Percentage of proliferating cells(S phase in cell cycle)markedly shrank from 40.42%for MSCs to 6.76%for MSC-derived neuron.Additionally,Wnt/β-catenin target genesβ-catenin and c-Myc were decreasingly expressed.Conclusion:chemically induced trans-differentiation from MSC to neuron caused similar MET-featured alteration in gene expression and proliferation to known MET,which might be underlied by de-activation of Wnt/β-catenin pathway.更多还原

【Abstract】 Mesenchymal-epithelial transition(MET) occurs commonly in both physiological and pathological context.Objective:To observe MET-associated alteration during the trans-differentiation from MSCs to neuron-like cells,and to explore the possible molecular mechanism.Methods:Bone marrow MSCs were isolated from rat femur and purified in continuous cell culture.After induced differentiation to neuron-like cells by the combination of butylated hydroxyanisole(BHA)and dimethyl sulfoxide(DMSO),cells were tested by comparative polymerase chain reaction(PCR) for the relative expression of MET biomarkers and transcription factors,and for cell cycle by flow cytometry.Meanwhile,target genes of Wnt/β-catenin pathway were also analyzed by comparative PCR to determine the possible involvement.Results:in MSC-induced neuron-like cells,MET-associated transcription factors such as Snail,Slug,ZEB1,ZEB2,and Twist were significantly attenuated in expression level.Mesenchymal marker vimentin was increased.Membrane protein E-cad was slightly down-regulated,while N-cad level was marginally elevated.Percentage of proliferating cells(S phase in cell cycle) markedly shrank from 40.42% for MSCs to 6.76% for MSC-derived neuron.Additionally,Wnt/β-catenin target genes β-catenin and c-Myc were decreasingly expressed.Conclusion:chemically induced trans-differentiation from MSC to neuron caused similar MET-featured alteration in gene expression and proliferation to known MET,which might be underlied by de-activation of Wnt/ β-catenin pathway.更多还原