Escherichia coli O157:H7 is one of the major foodborne pathogenic bacterial that cause infectious diseases in humans.The previous found that a combination of kojic acid and tea polyphenols exhibited better activity ag...Escherichia coli O157:H7 is one of the major foodborne pathogenic bacterial that cause infectious diseases in humans.The previous found that a combination of kojic acid and tea polyphenols exhibited better activity against E.coli O157:H7 than using either alone.This study aimed to explore responses underlying the antibacterial mechanisms of kojic acid and tea polyphenols from the gene level.The functional enrichment analysis by comparing kojic acid and tea polyphenols individually or synergistically against E.coli O157:H7 found that acid resistance systems in kojic acid were activated,and the cell membrane and genomic DNA were destructed in the cells,resulting in“oxygen starvation”.The oxidative stress response triggered by tea polyphenols inhibited both sulfur uptake and the synthesis of ATP,which affected the bacteria's life metabolic process.Interestingly,we found that kojic acid combined with tea polyphenols hindered the uptake of iron that played an essential role in the synthesis of DNA,respiration,tricarboxylic acid cycle.The results suggested that the iron uptake pathways may represent a novel approach for kojic acid and tea polyphenols synergistically against E.coli O157:H7 and provided a theoretical basis for bacterial pathogen control in the food industry.展开更多
目的探讨环指蛋白157(RING finger protein 157,RNF157)在黑素瘤细胞增殖中的作用及相关机制。方法采用慢病毒转染技术构建RNF157过表达黑素瘤细胞株,小干扰RNA(siRNA)技术构建敲减RNF157的黑素瘤细胞株,采用实时荧光定量聚合酶链反应(q...目的探讨环指蛋白157(RING finger protein 157,RNF157)在黑素瘤细胞增殖中的作用及相关机制。方法采用慢病毒转染技术构建RNF157过表达黑素瘤细胞株,小干扰RNA(siRNA)技术构建敲减RNF157的黑素瘤细胞株,采用实时荧光定量聚合酶链反应(qRT-PCR)和Western印迹实验验证转染效果。采用MTT实验、克隆形成实验检测RNF157过表达黑素瘤细胞和敲减RNF157黑素瘤细胞的细胞增殖能力。采用裸鼠皮下成瘤实验及免疫组化染色检测RNF157过表达黑素瘤细胞体外增殖能力。采用免疫共沉淀(Co-IP)结合质谱分析、蛋白质组学和泛素化组学,探究RNF157在ERK通路中的潜在作用机制。结果免疫组化染色结果显示,RNF157在黑素瘤组织中的表达高于瘤旁组织(P<0.01)。qRT-PCR和Western印迹实验结果显示,过表达组的RNF157表达水平显著高于对照组(P<0.0001);si3转染效率最高,si3组的RNF157表达水平显著低于对照组(P<0.01)。MTT实验、克隆形成实验结果显示,RNF157过表达可促进黑素瘤细胞增殖,敲减RNF157抑制黑素瘤细胞增殖。进一步检测发现,RNF157过表达可以激活黑素瘤细胞的ERK通路。Co-IP结合质谱分析、蛋白质组学和泛素化组学结果显示,RNF157可结合小凹蛋白-1(caveolin-1,CAV1)并下调其表达水平,从而调控ERK通路。结论RNF157可促进黑素瘤细胞的体内外增殖;在黑素瘤细胞中,RNF157可能通过结合并下调CAV1表达水平,参与细胞内ERK通路激活。展开更多
基金supported by National Natural Science Foundation of China(31972021)R&D Projects in Key Areas of Guangdong Province(2019B020212003)+4 种基金the Science and Technology Program of Guangzhou,China(202206010177)Guangdong key research and development program(2021B0202060001)Foshan and agricultural academy cooperation projectGuangdong Modern Agriculture project(2022KJ117)Aquatic Products Center Project of GAAS。
文摘Escherichia coli O157:H7 is one of the major foodborne pathogenic bacterial that cause infectious diseases in humans.The previous found that a combination of kojic acid and tea polyphenols exhibited better activity against E.coli O157:H7 than using either alone.This study aimed to explore responses underlying the antibacterial mechanisms of kojic acid and tea polyphenols from the gene level.The functional enrichment analysis by comparing kojic acid and tea polyphenols individually or synergistically against E.coli O157:H7 found that acid resistance systems in kojic acid were activated,and the cell membrane and genomic DNA were destructed in the cells,resulting in“oxygen starvation”.The oxidative stress response triggered by tea polyphenols inhibited both sulfur uptake and the synthesis of ATP,which affected the bacteria's life metabolic process.Interestingly,we found that kojic acid combined with tea polyphenols hindered the uptake of iron that played an essential role in the synthesis of DNA,respiration,tricarboxylic acid cycle.The results suggested that the iron uptake pathways may represent a novel approach for kojic acid and tea polyphenols synergistically against E.coli O157:H7 and provided a theoretical basis for bacterial pathogen control in the food industry.
文摘目的探讨环指蛋白157(RING finger protein 157,RNF157)在黑素瘤细胞增殖中的作用及相关机制。方法采用慢病毒转染技术构建RNF157过表达黑素瘤细胞株,小干扰RNA(siRNA)技术构建敲减RNF157的黑素瘤细胞株,采用实时荧光定量聚合酶链反应(qRT-PCR)和Western印迹实验验证转染效果。采用MTT实验、克隆形成实验检测RNF157过表达黑素瘤细胞和敲减RNF157黑素瘤细胞的细胞增殖能力。采用裸鼠皮下成瘤实验及免疫组化染色检测RNF157过表达黑素瘤细胞体外增殖能力。采用免疫共沉淀(Co-IP)结合质谱分析、蛋白质组学和泛素化组学,探究RNF157在ERK通路中的潜在作用机制。结果免疫组化染色结果显示,RNF157在黑素瘤组织中的表达高于瘤旁组织(P<0.01)。qRT-PCR和Western印迹实验结果显示,过表达组的RNF157表达水平显著高于对照组(P<0.0001);si3转染效率最高,si3组的RNF157表达水平显著低于对照组(P<0.01)。MTT实验、克隆形成实验结果显示,RNF157过表达可促进黑素瘤细胞增殖,敲减RNF157抑制黑素瘤细胞增殖。进一步检测发现,RNF157过表达可以激活黑素瘤细胞的ERK通路。Co-IP结合质谱分析、蛋白质组学和泛素化组学结果显示,RNF157可结合小凹蛋白-1(caveolin-1,CAV1)并下调其表达水平,从而调控ERK通路。结论RNF157可促进黑素瘤细胞的体内外增殖;在黑素瘤细胞中,RNF157可能通过结合并下调CAV1表达水平,参与细胞内ERK通路激活。
