The key regulators and regeneration-associated genes involved in axonal regeneration of neurons after injury have not been clarified.In high-throughput sequencing,various factors influence the final sequencing results...The key regulators and regeneration-associated genes involved in axonal regeneration of neurons after injury have not been clarified.In high-throughput sequencing,various factors influence the final sequencing results,including the number and size of cells,the depth of sequencing,and the method of cell separation.There is still a lack of research on the detailed molecular expression profile during the regeneration of dorsal root ganglion neuron axon.In this study,we performed lase r-capture microdissection coupled with RNA sequencing on dorsal root ganglion neurons at 0,3,6,and 12 hours and 1,3,and 7 days after sciatic nerve crush in rats.We identified three stages after dorsal root ganglion injury:early(3-12 hours),pre-regeneration(1 day),and regeneration(3-7 days).Gene expression patterns and related function enrichment res ults showed that one module of genes was highly related to axonal regeneration.We verified the up-regulation of activating transcription factor 3(Atf3),Kruppel like factor 6(Klf6),AT-rich inte raction domain 5A(Arid5α),CAMP responsive element modulator(Crem),and FOS like 1,AP-1 transcription factor Subunit(Fosl1) in dorsal root ganglion neurons after injury.Suppressing these transcription factors(Crem,Arid5o,Fosl1 and Klf6) reduced axonal regrowth in vitro.As the hub transcription factor,Atf3 showed higher expression and activity at the preregeneration and regeneration stages.G protein-coupled estrogen receptor 1(Gper1),inte rleukin 12a(Il12α),estrogen receptor 1(ESR1),and interleukin 6(IL6) may be upstream factors that trigger the activation of Atf3 during the repair of axon injury in the early stage.Our study presents the detailed molecular expression profile during axonal regeneration of dorsal root ganglion neurons after peripheral nerve injury.These findings may provide reference for the clinical screening of molecular targets for the treatment of peripheral nerve injury.展开更多
Background:Nucleolar protein 6(NOL6)is a nucleolar RNA-associated protein that is highly conserved between species.It has been proved to be associated with the prognosis of liver cancer.However,the underlying mechanis...Background:Nucleolar protein 6(NOL6)is a nucleolar RNA-associated protein that is highly conserved between species.It has been proved to be associated with the prognosis of liver cancer.However,the underlying mechanism has not been fully established.This study aimed to assess the relationship between NOL6 and liver cancer prognosis.Methods:We constructed anNOL6-short hairpin RNA(shRNA)-expressing lentivirus.Through viral transfection,cell growth assay and fluorescence-activated cell sorting,we evaluated the effect of shRNA-mediatedNOL6 knockdown on the proliferation,colony formation,and apoptosis of hepatocellular carcinoma(HCC)cells.The relationship betweenNOL6 expression and HCC patient survival has been established through bioinformatics analysis.We also explored the downstream molecular regulatory network ofNOL6 in HCC by performing an Ingenuity Pathway Analysis in the database.Results:IncreasedNOL6 expression was detected in HCC cells compared to normal controls;HCC patients with highNOL6 expression had poorer prognoses than those with low expression.NOL6 knockdown inhibited HCC cell proliferation,apoptosis,and colony formation.Also,MAPK8,CEBPA,andFOSL1 were selected as potential downstream genes ofNOL6.Conclusions:NOL6 up-regulates HCC cell proliferation and affects downstream expression of related genes.Moreover,NOL6 is considered to be associated with poor prognosis in HCC patients.展开更多
基金supported by the National Natural Science Foundation of China,Nos. 31730031 and 32130060the National Major Project of Research and Development,No. 2017YFA0104700the Natural Science Foundation of Jiangsu Province,No. BK20202013 (all to XSG)。
文摘The key regulators and regeneration-associated genes involved in axonal regeneration of neurons after injury have not been clarified.In high-throughput sequencing,various factors influence the final sequencing results,including the number and size of cells,the depth of sequencing,and the method of cell separation.There is still a lack of research on the detailed molecular expression profile during the regeneration of dorsal root ganglion neuron axon.In this study,we performed lase r-capture microdissection coupled with RNA sequencing on dorsal root ganglion neurons at 0,3,6,and 12 hours and 1,3,and 7 days after sciatic nerve crush in rats.We identified three stages after dorsal root ganglion injury:early(3-12 hours),pre-regeneration(1 day),and regeneration(3-7 days).Gene expression patterns and related function enrichment res ults showed that one module of genes was highly related to axonal regeneration.We verified the up-regulation of activating transcription factor 3(Atf3),Kruppel like factor 6(Klf6),AT-rich inte raction domain 5A(Arid5α),CAMP responsive element modulator(Crem),and FOS like 1,AP-1 transcription factor Subunit(Fosl1) in dorsal root ganglion neurons after injury.Suppressing these transcription factors(Crem,Arid5o,Fosl1 and Klf6) reduced axonal regrowth in vitro.As the hub transcription factor,Atf3 showed higher expression and activity at the preregeneration and regeneration stages.G protein-coupled estrogen receptor 1(Gper1),inte rleukin 12a(Il12α),estrogen receptor 1(ESR1),and interleukin 6(IL6) may be upstream factors that trigger the activation of Atf3 during the repair of axon injury in the early stage.Our study presents the detailed molecular expression profile during axonal regeneration of dorsal root ganglion neurons after peripheral nerve injury.These findings may provide reference for the clinical screening of molecular targets for the treatment of peripheral nerve injury.
基金supported by a grant from the Natural Science Foundation(No.81370069)。
文摘Background:Nucleolar protein 6(NOL6)is a nucleolar RNA-associated protein that is highly conserved between species.It has been proved to be associated with the prognosis of liver cancer.However,the underlying mechanism has not been fully established.This study aimed to assess the relationship between NOL6 and liver cancer prognosis.Methods:We constructed anNOL6-short hairpin RNA(shRNA)-expressing lentivirus.Through viral transfection,cell growth assay and fluorescence-activated cell sorting,we evaluated the effect of shRNA-mediatedNOL6 knockdown on the proliferation,colony formation,and apoptosis of hepatocellular carcinoma(HCC)cells.The relationship betweenNOL6 expression and HCC patient survival has been established through bioinformatics analysis.We also explored the downstream molecular regulatory network ofNOL6 in HCC by performing an Ingenuity Pathway Analysis in the database.Results:IncreasedNOL6 expression was detected in HCC cells compared to normal controls;HCC patients with highNOL6 expression had poorer prognoses than those with low expression.NOL6 knockdown inhibited HCC cell proliferation,apoptosis,and colony formation.Also,MAPK8,CEBPA,andFOSL1 were selected as potential downstream genes ofNOL6.Conclusions:NOL6 up-regulates HCC cell proliferation and affects downstream expression of related genes.Moreover,NOL6 is considered to be associated with poor prognosis in HCC patients.
