Kruppel-like factor 4（Klf4） is a zinc finger transcription factor and plays crucial roles in Xenopus embryogenesis.However, its regulation during embryogenesis is still unclear. Here, we report that Tcf711, a key do...Kruppel-like factor 4（Klf4） is a zinc finger transcription factor and plays crucial roles in Xenopus embryogenesis.However, its regulation during embryogenesis is still unclear. Here, we report that Tcf711, a key downstream transducer of the Wnt signaling pathway, could promote Klf4 transcription and stimulate Klf4 promoter activity in early Xenopus embryos. Furthermore, cycloheximide treatment showed a direct effect on Klf4 transcription facilitated by Tcf711. Moreover, the dominant negative form of Tcf711（dnTcf711）, which lacks N-terminus of the β-catenin binding motif, could still activate Klf4 transcription, suggesting that this regulation is Wnt/β-catenin independent.Taken together, our results demonstrate that Tcf711 lies upstream of Klf4 to maintain its expression level during Xenopus embryogenesis.展开更多
Quorum sensing(Qs)inhibition has emerged as a promising target for directed drug design,providing an appealing strategy for developing antimicrobials,particularly against infections caused by drug-resistant pathogens....Quorum sensing(Qs)inhibition has emerged as a promising target for directed drug design,providing an appealing strategy for developing antimicrobials,particularly against infections caused by drug-resistant pathogens.In this study,we designed and synthesized a total of 33β-nitrostyrene derivatives using 1-nitro-2-phenylethane(NPe)as the lead compound,to target the facultative anaerobic bacterial pathogen Serratia marcescens.The QS-inhibitory effects of these compounds were evaluated using S.marcescens NJ01 and the reporter strain Chromobacterium violaceum CV026.Among the 33 newβ-nitrostyrene derivatives,(E)-1-methyl-4-(2-nitrovinyl)benzene(m-NPe,compound 28)was proven to be a potent inhibitor that reduced biofilm formation of S.marcescens NJ01 by 79%.Scanning electron microscopy(SEM)and confocal laser scanning microscopy(CLSM)results revealed that treatment with m-NPe(50μg/ml)not only enhanced the susceptibility of the formed biofilms but also disrupted the architecture of biofilms by 84%.m-NPe(50μg/ml)decreased virulence factors in S.marcescens NJ01,reducing the activity of protease,prodigiosin,and extracellular polysaccharide(EPs)by 36%,72%,and 52%,respectively.In S.marcescens 4547,the activities of hemolysin and EPs were reduced by 28%and 40%,respectively,outperforming the positive control,vanillic acid(VAN).The study also found that the expression levels of QS-and biofilm-related genes(flhD,fimA,fimC,sodB,bsmB,pigA,pigC,and shlA)were downregulated by 1.21-to 2.32-fold.Molecular dynamics analysis showed that m-NPe could bind stably to SmaR,Rhll,RhiR,LasR,and CviR proteins in a 0.1 M sodium chloride solution.Importantly,a microscale thermophoresis(MST)test revealed that SmaR could be a target protein for the screening of a quorum sensing inhibitor(QSl)against S.marcescens.Overall,this study highlights the efficacy of m-NPe in suppressing the virulence factors of S.marcescens,identifying it as a new potential Qsl and antibiofilm agent capable of restoring or improving antimicrobial drug sensitivity.展开更多
目的探讨人肝细胞癌中核转录因子Kruppel样因子4(KLF4)与叉头框蛋白M1(FOXM1)基因表达相关性,以及两种转录因子表达对肝细胞癌生长和增殖的影响。方法采用实时荧光定量PCR技术检测50例肝细胞癌患者肝癌组织及8株肝癌细胞中KLF4m RNA和FO...目的探讨人肝细胞癌中核转录因子Kruppel样因子4(KLF4)与叉头框蛋白M1(FOXM1)基因表达相关性,以及两种转录因子表达对肝细胞癌生长和增殖的影响。方法采用实时荧光定量PCR技术检测50例肝细胞癌患者肝癌组织及8株肝癌细胞中KLF4m RNA和FOXM1 m RNA表达水平并分析其相关性;KLF4过表达质粒和KLF4 si RNA分别转染人肝癌细胞Hep3B和BEL-7402,采用实时荧光定量PCR、蛋白印迹实验检测KLF4 m RNA、FOXM1m RNA及KLF4蛋白、FOXM1蛋白表达水平;通过CCK8实验检测KLF4和FOXM1基因对细胞增殖的影响。结果肝癌组织和细胞中KLF4低表达,而FOXM1高表达,两者呈负相关(r=-0.462,P=0.001);KLF4基因过表达抑制肝癌细胞增殖,而FOXM1基因过表达促进肝癌细胞增殖,FOXM1基因过表达能恢复KLF4对肝癌细胞抑制增殖作用。结论在肝细胞癌中,KLF4和FOXM1基因表达呈负相关,两者不同的表达水平可影响肝癌细胞的增殖。展开更多
基金supported by the Start-up Funding of Henan University of Science and Technology(13480027) to Q. C.the Key Science Foundation of Nanjing Medical University(2015NJMUZD002)+2 种基金the Natural Science Foundation of Higher Education Institutions of Jiangsu Province(16KJB-180020)Natural Science Foundation of Jiangsu Province (BK20171053)National Natural Science Funds of China (81702747) to C.L
文摘Kruppel-like factor 4（Klf4） is a zinc finger transcription factor and plays crucial roles in Xenopus embryogenesis.However, its regulation during embryogenesis is still unclear. Here, we report that Tcf711, a key downstream transducer of the Wnt signaling pathway, could promote Klf4 transcription and stimulate Klf4 promoter activity in early Xenopus embryos. Furthermore, cycloheximide treatment showed a direct effect on Klf4 transcription facilitated by Tcf711. Moreover, the dominant negative form of Tcf711（dnTcf711）, which lacks N-terminus of the β-catenin binding motif, could still activate Klf4 transcription, suggesting that this regulation is Wnt/β-catenin independent.Taken together, our results demonstrate that Tcf711 lies upstream of Klf4 to maintain its expression level during Xenopus embryogenesis.
基金This work was supported by the National Natural Science Foundation of China(Nos.82160664 and 31760246)the Hainan Province Science and Technology Special Fund(ZDYF2024SHFZ103)+1 种基金the Natural Science Foundation of Hainan Province(222RC557 and 221QN170)Anhui Laboratory of Molecule-Based Materials(fzj21006).
文摘Quorum sensing(Qs)inhibition has emerged as a promising target for directed drug design,providing an appealing strategy for developing antimicrobials,particularly against infections caused by drug-resistant pathogens.In this study,we designed and synthesized a total of 33β-nitrostyrene derivatives using 1-nitro-2-phenylethane(NPe)as the lead compound,to target the facultative anaerobic bacterial pathogen Serratia marcescens.The QS-inhibitory effects of these compounds were evaluated using S.marcescens NJ01 and the reporter strain Chromobacterium violaceum CV026.Among the 33 newβ-nitrostyrene derivatives,(E)-1-methyl-4-(2-nitrovinyl)benzene(m-NPe,compound 28)was proven to be a potent inhibitor that reduced biofilm formation of S.marcescens NJ01 by 79%.Scanning electron microscopy(SEM)and confocal laser scanning microscopy(CLSM)results revealed that treatment with m-NPe(50μg/ml)not only enhanced the susceptibility of the formed biofilms but also disrupted the architecture of biofilms by 84%.m-NPe(50μg/ml)decreased virulence factors in S.marcescens NJ01,reducing the activity of protease,prodigiosin,and extracellular polysaccharide(EPs)by 36%,72%,and 52%,respectively.In S.marcescens 4547,the activities of hemolysin and EPs were reduced by 28%and 40%,respectively,outperforming the positive control,vanillic acid(VAN).The study also found that the expression levels of QS-and biofilm-related genes(flhD,fimA,fimC,sodB,bsmB,pigA,pigC,and shlA)were downregulated by 1.21-to 2.32-fold.Molecular dynamics analysis showed that m-NPe could bind stably to SmaR,Rhll,RhiR,LasR,and CviR proteins in a 0.1 M sodium chloride solution.Importantly,a microscale thermophoresis(MST)test revealed that SmaR could be a target protein for the screening of a quorum sensing inhibitor(QSl)against S.marcescens.Overall,this study highlights the efficacy of m-NPe in suppressing the virulence factors of S.marcescens,identifying it as a new potential Qsl and antibiofilm agent capable of restoring or improving antimicrobial drug sensitivity.
文摘目的探讨人肝细胞癌中核转录因子Kruppel样因子4(KLF4)与叉头框蛋白M1(FOXM1)基因表达相关性,以及两种转录因子表达对肝细胞癌生长和增殖的影响。方法采用实时荧光定量PCR技术检测50例肝细胞癌患者肝癌组织及8株肝癌细胞中KLF4m RNA和FOXM1 m RNA表达水平并分析其相关性;KLF4过表达质粒和KLF4 si RNA分别转染人肝癌细胞Hep3B和BEL-7402,采用实时荧光定量PCR、蛋白印迹实验检测KLF4 m RNA、FOXM1m RNA及KLF4蛋白、FOXM1蛋白表达水平;通过CCK8实验检测KLF4和FOXM1基因对细胞增殖的影响。结果肝癌组织和细胞中KLF4低表达,而FOXM1高表达,两者呈负相关(r=-0.462,P=0.001);KLF4基因过表达抑制肝癌细胞增殖,而FOXM1基因过表达促进肝癌细胞增殖,FOXM1基因过表达能恢复KLF4对肝癌细胞抑制增殖作用。结论在肝细胞癌中,KLF4和FOXM1基因表达呈负相关,两者不同的表达水平可影响肝癌细胞的增殖。
