Background：Abnormal neuronal differentiation plays an important role in central nervous system （CNS） development abnormalities such as Down syndrome （DS）,a disorder that results directly from overexpression of ge...Background：Abnormal neuronal differentiation plays an important role in central nervous system （CNS） development abnormalities such as Down syndrome （DS）,a disorder that results directly from overexpression of genes in trisomic cells.Receptor-interacting protein 140 （RIP 140） is significantly upregulated in DS brains,suggesting its involvement in DS CNS development abnormalities.However,the role of RIP140 in neuronal differentiation is still not clear.The current study aimed to investigate the effect of RIP 140 overexpression on the differentiation of neuro-2a （N2a） neuroblastoma cells,in vitro.Methods：Stably RIP 140-overexpressing N2a （N2a-RIP140） cells were used as a neurodevelopmental model,and were constructed by lipofection and overexpression validated by real-time polymerase chain reaction and Western blot.Retinoic acid （RA） was used to stimulate N2a differentiation.Combining the expression of Tuj 1 at the mRNA and protein levels,the percentage of cells baring neurites,and the number of neurites per cell body was semi-quantified to determine the effect of RIP 140 on differentiation of N2a cells.Furthermore,western blot and the ERK1/2 inhibitor U0126 were used to identify the specific signaling pathway by which RIP 140 induces differentiation of N2a cells.Statistical significance of the differences between groups was determined by one-way analysis of variance followed by the Dunnett test.Results：Compared to untransfected N2a cells RIP140 expression in N2a-RIP 140 cells was remarkably upregulated at both the mRNA and protein levels.N2a-RIP 140 cells had a significantly increased percentage of cells baring neurites,and numbers of neurites per cell,as compared to N2a cells,in the absence and presence of RA （P 〈 0.05）.In addition,Tuj l,a neuronal biomarker,was strongly upregulated in N2a-RIP140 cells （P 〈 0.05） and phosphorylated ERK1/2 （p-ERK1/2） levels in N2a-RIP140 cells were dramatically increased,while differentiation was inhibited by the ERK 1/2-specific inhibitor U0126.Conclusions：RIP140 overexpression promotes N2a cell neuronal differentiation by activating the ERK1/2 pathway.展开更多
基金This research was supported by the grants from the National Natural Science Foundation of China （No. 30872792） and Chief Funds of the National Natural Science Foundation of China （No. 31340024）.
文摘Background：Abnormal neuronal differentiation plays an important role in central nervous system （CNS） development abnormalities such as Down syndrome （DS）,a disorder that results directly from overexpression of genes in trisomic cells.Receptor-interacting protein 140 （RIP 140） is significantly upregulated in DS brains,suggesting its involvement in DS CNS development abnormalities.However,the role of RIP140 in neuronal differentiation is still not clear.The current study aimed to investigate the effect of RIP 140 overexpression on the differentiation of neuro-2a （N2a） neuroblastoma cells,in vitro.Methods：Stably RIP 140-overexpressing N2a （N2a-RIP140） cells were used as a neurodevelopmental model,and were constructed by lipofection and overexpression validated by real-time polymerase chain reaction and Western blot.Retinoic acid （RA） was used to stimulate N2a differentiation.Combining the expression of Tuj 1 at the mRNA and protein levels,the percentage of cells baring neurites,and the number of neurites per cell body was semi-quantified to determine the effect of RIP 140 on differentiation of N2a cells.Furthermore,western blot and the ERK1/2 inhibitor U0126 were used to identify the specific signaling pathway by which RIP 140 induces differentiation of N2a cells.Statistical significance of the differences between groups was determined by one-way analysis of variance followed by the Dunnett test.Results：Compared to untransfected N2a cells RIP140 expression in N2a-RIP 140 cells was remarkably upregulated at both the mRNA and protein levels.N2a-RIP 140 cells had a significantly increased percentage of cells baring neurites,and numbers of neurites per cell,as compared to N2a cells,in the absence and presence of RA （P 〈 0.05）.In addition,Tuj l,a neuronal biomarker,was strongly upregulated in N2a-RIP140 cells （P 〈 0.05） and phosphorylated ERK1/2 （p-ERK1/2） levels in N2a-RIP140 cells were dramatically increased,while differentiation was inhibited by the ERK 1/2-specific inhibitor U0126.Conclusions：RIP140 overexpression promotes N2a cell neuronal differentiation by activating the ERK1/2 pathway.
