Objective To investigate the regulatory relationship of Protein Phosphatase 2 Regulatory Subunit B"Alpha(PPP2R3A)and hexokinase 1(HK1)in glycolysis of hepatocellular carcinoma(HCC).Methods In HepG2 and Huh7 cells...Objective To investigate the regulatory relationship of Protein Phosphatase 2 Regulatory Subunit B"Alpha(PPP2R3A)and hexokinase 1(HK1)in glycolysis of hepatocellular carcinoma(HCC).Methods In HepG2 and Huh7 cells,PPP2R3A expression was silenced by small interfering RNA(siRNA)and overexpression by plasmid transfection.The PPP2R3A-related genes were searched by RNA sequencing.Glycolysis levels were measured by glucose uptake and lactate production.QRT-PCR,ELISA,western blot and immunofluorescence assay were performed to detect the changes of PPP2R3A and HK1.Cell proliferation,migration and invasion assay were used to study the roles of HK1 regulation by PPP2R3A.Results RNA sequencing data revealed that PPP2R3A siRNA significantly downregulated the expression of HK1.PPP2R3A gene overexpression promotes,while gene silencing suppresses,the level of HK1 and glycolysis in HCC cells.In HCC tissue samples,PPP2R3A and HK1 were colocalized in the cytoplasm,and their expression showed a positive correlation.HK1 inhibition abrogated the promotion of glycolysis,proliferation,migration and invasion by PPP2R3A overexpression in liver cancer cells.Conclusion Our findings showed the correlation of PPP2R3A and HK1 in the glycolysis of HCC,which reveals a new mechanism for the oncogenic roles of PPP2R3A in cancer.展开更多
Objective To detect the relationship between the polymorphism of the glycogen-targeting regulatory subunit of the skeletal muscle glycogen-associated protein phosphatase 1 (PPP1R3) gene and type 2 diabetes by case-con...Objective To detect the relationship between the polymorphism of the glycogen-targeting regulatory subunit of the skeletal muscle glycogen-associated protein phosphatase 1 (PPP1R3) gene and type 2 diabetes by case-control study. Methods We genotyped the PPP1R3 gene Asp905Tyr polymorphism and a common 3'-untranslated region AT (AU)-rich element (ARE) polymorphism in 101 type 2 diabetic patients and 101controls by oligonucleotide ligation assay (OLA) and polyacrylamide gel elecrophoresis, respectively. Results Subjects with Tyr/Tyr genotypes whose body mass index (BMI)<25 were used as the reference group. Those whose BMI25 with Asp905 had a 3.66-fold increase (95% CI: 1.48-9.06, P=0.005) in type 2 diabetes risk. No association was found between 3'UTR ARE polymorphism and type 2 diabetes mellitus (OR=1.15; 95% CI: 0.62-2.14, P=0.65). Conclusion A joint effect between the Asp905 and BMI increases the risk of type 2 diabetes, and Asp905Tyr and ARE polymorphism of PPP1R3 gene are not the major diabetogenic gene variants in Chinese population.展开更多
After leaving the testis,mammalian sperm undergo a sequential maturation process in the epididymis followed by capacitation during their movement through the female reproductive tract.These phenotypic changes are asso...After leaving the testis,mammalian sperm undergo a sequential maturation process in the epididymis followed by capacitation during their movement through the female reproductive tract.These phenotypic changes are associated with modification of protein phosphorylation and membrane remodeling,which is requisite for sperm to acquire forward motility and induce fertilization.However,the molecular mechanisms underlying sperm maturation and capacitation are still not fully understood.Herein,we show that PPP3R2,a testis-specific regulatory subunit of protein phosphatase 3(an isoform of calcineurin in the testis),is essential for sperm maturation and capacitation.Knockout of Ppp3r2 in mice leads to male sterility due to sperm motility impairment and morphological defects.One very noteworthy change includes increases in sperm membrane stiffness.Moreover,PPP3R2 regulates sperm maturation and capacitation via(i)modulation of membrane diffusion barrier function at the annulus and(ii)facilitation of cholesterol efflux during sperm capacitation.Taken together,PPP3R2 plays a critical role in modulating cholesterol efflux and mediating the dynamic control of membrane remodeling during sperm maturation and capacitation.展开更多
基金supported by National Natural Science Foundation of China [81372595]
文摘Objective To investigate the regulatory relationship of Protein Phosphatase 2 Regulatory Subunit B"Alpha(PPP2R3A)and hexokinase 1(HK1)in glycolysis of hepatocellular carcinoma(HCC).Methods In HepG2 and Huh7 cells,PPP2R3A expression was silenced by small interfering RNA(siRNA)and overexpression by plasmid transfection.The PPP2R3A-related genes were searched by RNA sequencing.Glycolysis levels were measured by glucose uptake and lactate production.QRT-PCR,ELISA,western blot and immunofluorescence assay were performed to detect the changes of PPP2R3A and HK1.Cell proliferation,migration and invasion assay were used to study the roles of HK1 regulation by PPP2R3A.Results RNA sequencing data revealed that PPP2R3A siRNA significantly downregulated the expression of HK1.PPP2R3A gene overexpression promotes,while gene silencing suppresses,the level of HK1 and glycolysis in HCC cells.In HCC tissue samples,PPP2R3A and HK1 were colocalized in the cytoplasm,and their expression showed a positive correlation.HK1 inhibition abrogated the promotion of glycolysis,proliferation,migration and invasion by PPP2R3A overexpression in liver cancer cells.Conclusion Our findings showed the correlation of PPP2R3A and HK1 in the glycolysis of HCC,which reveals a new mechanism for the oncogenic roles of PPP2R3A in cancer.
文摘Objective To detect the relationship between the polymorphism of the glycogen-targeting regulatory subunit of the skeletal muscle glycogen-associated protein phosphatase 1 (PPP1R3) gene and type 2 diabetes by case-control study. Methods We genotyped the PPP1R3 gene Asp905Tyr polymorphism and a common 3'-untranslated region AT (AU)-rich element (ARE) polymorphism in 101 type 2 diabetic patients and 101controls by oligonucleotide ligation assay (OLA) and polyacrylamide gel elecrophoresis, respectively. Results Subjects with Tyr/Tyr genotypes whose body mass index (BMI)<25 were used as the reference group. Those whose BMI25 with Asp905 had a 3.66-fold increase (95% CI: 1.48-9.06, P=0.005) in type 2 diabetes risk. No association was found between 3'UTR ARE polymorphism and type 2 diabetes mellitus (OR=1.15; 95% CI: 0.62-2.14, P=0.65). Conclusion A joint effect between the Asp905 and BMI increases the risk of type 2 diabetes, and Asp905Tyr and ARE polymorphism of PPP1R3 gene are not the major diabetogenic gene variants in Chinese population.
基金supported by grants from the National Natural Science Foundation of China(81571487,81701503,and 81971437)Science and Technology Commission of Shanghai Municipality(16ZR1418600).
文摘After leaving the testis,mammalian sperm undergo a sequential maturation process in the epididymis followed by capacitation during their movement through the female reproductive tract.These phenotypic changes are associated with modification of protein phosphorylation and membrane remodeling,which is requisite for sperm to acquire forward motility and induce fertilization.However,the molecular mechanisms underlying sperm maturation and capacitation are still not fully understood.Herein,we show that PPP3R2,a testis-specific regulatory subunit of protein phosphatase 3(an isoform of calcineurin in the testis),is essential for sperm maturation and capacitation.Knockout of Ppp3r2 in mice leads to male sterility due to sperm motility impairment and morphological defects.One very noteworthy change includes increases in sperm membrane stiffness.Moreover,PPP3R2 regulates sperm maturation and capacitation via(i)modulation of membrane diffusion barrier function at the annulus and(ii)facilitation of cholesterol efflux during sperm capacitation.Taken together,PPP3R2 plays a critical role in modulating cholesterol efflux and mediating the dynamic control of membrane remodeling during sperm maturation and capacitation.
