Effect Of Kctd10 On Liver Stem Cell Marker Genes,immune Molecules And Notch Signaling Pathway In Vivo

Objective:Studying the effect of Kctd10 deficiency on liver stem cell marker genes and immune molecules involved in vivo.Methods:The liver specific knockout mice were established by using the Cre/lox P recombinase system,PCR amplification-agarose gel electrophoresis was used to identify the mouse genotype.The knockout effect was confirmed by Western Blot analysis,fluorescence quantitative PCR and immune-histochemistry.HE staining was used to analyze the structure changes of the liver tissues in Alb cre⁺;Kctd10^flox/floxmouse.Western blot analysis,fluorescence quantitative PCR and immune-histochemistry were used to determine the expression of liver stem cells marker genes after Kctd10 deletion in mouse liver.Western Blot analysis and q PCR were used to determine the modulation of Notch1 and other Notch signaling pathway associated proteins.Western Blot analysis,real-time PCR analysis,and RNA sequencing to check the changes in Notch signaling related genes and immune molecules after specifically knocking out the Kctd10 gene in mouse liver.Results:（1）After specific knockout of Kctd10 gene in the liver of mice,the m RNA level and protein level of Kctd10 were significantly reduced compared with the wild-type mice in the same litter.（2）After specific knockout of liver Kctd10 gene in mice,the expression of liver stem cell-related marker genes（Lgr5,Ck7,Ck19）was up-regulated compared with that of wild-type mice among the same littermates.（3）After specific knockout of mouse liver Kctd10 gene,the protein levels of Notch1protein and some key members of Notch were decreased compared with that in the wild-type mice.（4）The m RNA levels of Th1/Th2 immune molecules（H2-aa,H2-ab1,H2-eb1,H2-dma,H2-dmb1）and its related regulators（Rorc,Stat1,Zap70）were decreased after specific knockout of Kctd10 gene in mouse liver.Conclusion:1.Liver specific deletion of Kctd10 up-regulates the expression of liver stem cell marker genes（Lgr5,Ck7,Ck19）;2.Liver specific deletion of Kctd10 down-regulates the expression of Th1/Th2 immune molecules（H2-aa,H2-ab1,H2-eb1,H2-dma,H2-dmb1）and immune regulators（Rorc,Stat1,Zap70）;3.Loss of Kctd10 liver specificity leads to disorder of Notch signaling pathway（ligand level decreased）;4.The changes in the expression profile of liver stem cells and Th1/Th2immuno-molecular expression profile of Kctd10 liver-specific deletion may be related to Notch signaling pathway.