BACKGROUND Sarcosine dehydrogenase(SARDH)and C-X-C motif chemokine ligand 1(CXCL1)have been identified as potential tumor regulators,with growing evidence linking them to cancer progression.However,their specific role...BACKGROUND Sarcosine dehydrogenase(SARDH)and C-X-C motif chemokine ligand 1(CXCL1)have been identified as potential tumor regulators,with growing evidence linking them to cancer progression.However,their specific roles,regulatory mechanisms,and influence on key signaling pathways remain unclear.AIM To investigate the regulatory mechanisms of SARDH and CXCL1 in cancer cells and their impact on key signaling pathways.METHODS Real-time quantitative polymerase chain reaction and western blot analyses were used to assess the expression levels of SARDH and CXCL1 and their effects on protein kinase B(Akt)and extracellular signal-regulated kinase(ERK)signaling pathways.Gene overexpression was induced using an expression vector,while gene silencing was achieved using short hairpin RNA and small interfering RNA.CCK-8,migration,and invasion assays were used to evaluate the impact of gene suppression and overexpression on cancer cell proliferation,migration,and invasion.RESULTS SARDH silencing significantly enhanced cancer cell proliferation,whereas its overexpression suppressed proliferation in the early stages of the experiment.CXCL1 silencing reduced cancer cell migration and invasion.SARDH overexpression inhibited cell migration,invasion,and adhesion while increasing apoptosis.Conversely,SARDH silencing reversed these effects.Furthermore,simultaneous silencing of SARDH and CXCL1 strongly activated the Akt and ERK signaling pathways,indicating the potential role of these pathways in regulating cellular functions influenced by these genes.CONCLUSION This study revealed that SARDH and CXCL1 regulate cancer cell growth,migration,and invasion through Akt and ERK signaling pathways,highlighting their potential as therapeutic targets for cancer treatment.展开更多
The title compound (C10H12N2O7, Mr = 272.22) crystallizes in triclinic, space group P1 with a = 5.532(2), b = 9.760(4), c = 11.731(5) ?, α = 68.107(7), β = 89.179(7), γ = 77.830(7)o, V = 573.1(4) ?3, Z = 2, Dc = 1....The title compound (C10H12N2O7, Mr = 272.22) crystallizes in triclinic, space group P1 with a = 5.532(2), b = 9.760(4), c = 11.731(5) ?, α = 68.107(7), β = 89.179(7), γ = 77.830(7)o, V = 573.1(4) ?3, Z = 2, Dc = 1.578 g/cm3, F(000) = 284 and μ(MoKa) = 0.136 mm-1. The final R = 0.0400 and wR = 0.0951 for 1468 observed reflections with I > 2σ(I). The title compound is a 1:1 adduct of sarcosine and 5-nitrosalicylic acid. The nitrogen atom of sarcosine is protonated, and the proton is from the carboxyl group of sarcosine and 5-nitrosalicylic acid with the probability of 50 percent for each. The 5-nitrosalicylic acid and sarcosine molecule of the title adduct are ABAB arranged along the c axis. There exist a lot of hydrogen bonds in the structure, linking sarcosine and 5-nitrosalicylic acid to form a three-dimensional network.展开更多
N-methyl-d-aspartic acid(NMDA),an amino acid existing in human and animal central nervous system,exerts agonist action on one of the glutamate receptor subtypes and is clinically used for treatment of diabetes,Parkin...N-methyl-d-aspartic acid(NMDA),an amino acid existing in human and animal central nervous system,exerts agonist action on one of the glutamate receptor subtypes and is clinically used for treatment of diabetes,Parkinson’s and Alzheimer’s syndrome.In this study,an enzymatic protocol for the chiral resolution of N-methyl-d,l-aspartic acid was built with a predicted N-demethylase(GenBank ID:OJV90073.1)from the genome of Chloroflexi bacterium 54-19.Through sequence alignment,the enzyme shares an identity of 32.19%to 2UZZ(PDB ID)with a conserved catalytic center.Recombinantly expressed in Bacillus subtilis WB600,the N-demethylase was characterized with optimal temperature and pH at 55℃and 7.5,and adaptive temperature and pH were 40-60℃ and 6-8.The effects of organic solvents and metal ions were investigated as well.Compared with other previously reported sarcosine oxidases,the enzyme showed a specific N-demethylation activity against N-methyl-l-aspartic acid according to the analysis by chiral liquid chromatography,LC-MS/MS and a detection by polarimeter.The results demonstrated 76%of 4.5 mM N-methyl-d,l-aspartic acid could be chirally separated by 7 mg·L^(−1) enzyme after reaction of 80 min.This work provided a foundation for mild synthesis of NMDA in industry.展开更多
基金Supported by the Qingpu Science and Technology Commission Project,No.QKY2022-13.
文摘BACKGROUND Sarcosine dehydrogenase(SARDH)and C-X-C motif chemokine ligand 1(CXCL1)have been identified as potential tumor regulators,with growing evidence linking them to cancer progression.However,their specific roles,regulatory mechanisms,and influence on key signaling pathways remain unclear.AIM To investigate the regulatory mechanisms of SARDH and CXCL1 in cancer cells and their impact on key signaling pathways.METHODS Real-time quantitative polymerase chain reaction and western blot analyses were used to assess the expression levels of SARDH and CXCL1 and their effects on protein kinase B(Akt)and extracellular signal-regulated kinase(ERK)signaling pathways.Gene overexpression was induced using an expression vector,while gene silencing was achieved using short hairpin RNA and small interfering RNA.CCK-8,migration,and invasion assays were used to evaluate the impact of gene suppression and overexpression on cancer cell proliferation,migration,and invasion.RESULTS SARDH silencing significantly enhanced cancer cell proliferation,whereas its overexpression suppressed proliferation in the early stages of the experiment.CXCL1 silencing reduced cancer cell migration and invasion.SARDH overexpression inhibited cell migration,invasion,and adhesion while increasing apoptosis.Conversely,SARDH silencing reversed these effects.Furthermore,simultaneous silencing of SARDH and CXCL1 strongly activated the Akt and ERK signaling pathways,indicating the potential role of these pathways in regulating cellular functions influenced by these genes.CONCLUSION This study revealed that SARDH and CXCL1 regulate cancer cell growth,migration,and invasion through Akt and ERK signaling pathways,highlighting their potential as therapeutic targets for cancer treatment.
文摘The title compound (C10H12N2O7, Mr = 272.22) crystallizes in triclinic, space group P1 with a = 5.532(2), b = 9.760(4), c = 11.731(5) ?, α = 68.107(7), β = 89.179(7), γ = 77.830(7)o, V = 573.1(4) ?3, Z = 2, Dc = 1.578 g/cm3, F(000) = 284 and μ(MoKa) = 0.136 mm-1. The final R = 0.0400 and wR = 0.0951 for 1468 observed reflections with I > 2σ(I). The title compound is a 1:1 adduct of sarcosine and 5-nitrosalicylic acid. The nitrogen atom of sarcosine is protonated, and the proton is from the carboxyl group of sarcosine and 5-nitrosalicylic acid with the probability of 50 percent for each. The 5-nitrosalicylic acid and sarcosine molecule of the title adduct are ABAB arranged along the c axis. There exist a lot of hydrogen bonds in the structure, linking sarcosine and 5-nitrosalicylic acid to form a three-dimensional network.
基金supported by the National Key Research and Development Program of China(2018YFA0900300,2021YFC2100300).
文摘N-methyl-d-aspartic acid(NMDA),an amino acid existing in human and animal central nervous system,exerts agonist action on one of the glutamate receptor subtypes and is clinically used for treatment of diabetes,Parkinson’s and Alzheimer’s syndrome.In this study,an enzymatic protocol for the chiral resolution of N-methyl-d,l-aspartic acid was built with a predicted N-demethylase(GenBank ID:OJV90073.1)from the genome of Chloroflexi bacterium 54-19.Through sequence alignment,the enzyme shares an identity of 32.19%to 2UZZ(PDB ID)with a conserved catalytic center.Recombinantly expressed in Bacillus subtilis WB600,the N-demethylase was characterized with optimal temperature and pH at 55℃and 7.5,and adaptive temperature and pH were 40-60℃ and 6-8.The effects of organic solvents and metal ions were investigated as well.Compared with other previously reported sarcosine oxidases,the enzyme showed a specific N-demethylation activity against N-methyl-l-aspartic acid according to the analysis by chiral liquid chromatography,LC-MS/MS and a detection by polarimeter.The results demonstrated 76%of 4.5 mM N-methyl-d,l-aspartic acid could be chirally separated by 7 mg·L^(−1) enzyme after reaction of 80 min.This work provided a foundation for mild synthesis of NMDA in industry.
