Multidrug Resistance Protein 2 (MRP2) is an ATP-dependent transmembrane protein that plays a pivotal role in the efflux of a wide variety of physiological substrates across the plasma membrane. Several studies have sh...Multidrug Resistance Protein 2 (MRP2) is an ATP-dependent transmembrane protein that plays a pivotal role in the efflux of a wide variety of physiological substrates across the plasma membrane. Several studies have shown that MRP2 can significantly affect the absorption, distribution, metabolism, excretion, and toxicity (ADMET) profiles of many therapeutic drugs and chemicals found in the environment and diet. This transporter can also efflux newly developed anticancer agents that target specific signaling pathways and are major clinical markers associated with multidrug resistance (MDR) of several types of cancers. MDR remains a major limitation to the advancement of the combinatorial chemotherapy regimen in cancer treatment. In addition to anticancer agents, MRP2 reduces the efficacy of various drug classes such as antivirals, antimalarials, and antibiotics. The unique role of MRP2 and its contribution to MDR makes it essential to profile drug-transporter interactions for all new and promising drugs. Thus, this current research seeks to identify modulators of MRP2 protein expression levels using cell-based assays. A unique recently approved FDA library (372 drugs) was screened using a high-throughput In-Cell ELISA assay to determine the effect of these therapeutic agents on protein expression levels of MRP2. A total of 49 FDA drugs altered MRP2 protein expression levels by more than 50% representing 13.17% of the compounds screened. Among the identified hits, thirty-nine (39) drugs increased protein expression levels whereas 10 drugs lowered protein expression levels of MRP2 after drug treatment. Our findings from this initial drug screening showed that modulators of MRP2 peregrinate multiple drug families and signify the importance of profiling drug interactions with this transporter. Data from this study provides essential information to improve combinatorial drug therapy and precision medicine as well as reduce the drug toxicity of various cancer chemotherapies.展开更多
目的探讨胆盐载体MRP1、MRP2在妊娠期肝内胆汁淤积症(ICP)胎盘的表达,分析胎盘胆盐载体与ICP发病的关系。方法收集8例正常早孕妇女绒毛组织、7例正常中孕妇女胎盘组织、20例正常晚孕妇女(对照组)胎盘组织,以及20例ICP患者胎盘组织,检测...目的探讨胆盐载体MRP1、MRP2在妊娠期肝内胆汁淤积症(ICP)胎盘的表达,分析胎盘胆盐载体与ICP发病的关系。方法收集8例正常早孕妇女绒毛组织、7例正常中孕妇女胎盘组织、20例正常晚孕妇女(对照组)胎盘组织,以及20例ICP患者胎盘组织,检测收集组织中胆盐载体MRP1和MRP2 mRNA的表达。结果ICP和正常妊娠各期胎盘组织中均有MRP2 mRNA的表达,而MRP1 mRNA在大部分标本上有表达;ICP组MRP1 mRNA和MRP2 mRNA表达量与对照组相比(99.94±73.17 vs 99.20±68.65;95.78±56.50 vs 142.20±91.27)差异无统计学意义(P>0.05);ICP组未用地塞米松治疗胎盘MRP2 mRNA表达量低于对照组(91.82±48.08 vs 142.20±91.27,P<0.05)。结论在未用地塞米松治疗的ICP患者胎盘组织中,MRP2 mRNA的表达量降低,这可能是引起ICP患者胎盘胆汁酸转运障碍,从而引起胎儿体内胆汁淤积的机制之一。展开更多
根据猪链球菌2型(Streptococcus suis type 2,SS2)溶菌酶释放蛋白(muramidase-released protein,MRP)和胞外蛋白因子(extracellular protein factor,EPF)的基因序列,各设计合成一对引物,以猪链球菌2型江苏分离株SS2-1的基因组为模板,扩...根据猪链球菌2型(Streptococcus suis type 2,SS2)溶菌酶释放蛋白(muramidase-released protein,MRP)和胞外蛋白因子(extracellular protein factor,EPF)的基因序列,各设计合成一对引物,以猪链球菌2型江苏分离株SS2-1的基因组为模板,扩增mrp基因1 801~2 513位序列和epf 基因1 783~2 563位序列,分别构建原核表达载体pET32a-mrp 、pET32a-epf,确定诱导表达的两种蛋白都具有免疫原性后,提取阳性克隆质粒各自进行双酶切并纯化,通过PCR串联两片段,将目的片段定向克隆到表达载体pET-32a中,重组质粒转化入大肠杆菌BL21,经IPTG诱导表达分子量约为74kD的融合蛋白.用制备的两种抗血清与纯化融合蛋白进行免疫转印,结果显示融合蛋白分别具有MRP与EPF的中和表位.用融合蛋白MRP-EPF免疫新西兰兔,以最小致死量猪链球菌强毒株SS2-1攻击,兔的存活率达50%(2/4),存活率明显高于单个表达产物.证实串联表达的融合蛋白为重要的保护性抗原.展开更多
文摘Multidrug Resistance Protein 2 (MRP2) is an ATP-dependent transmembrane protein that plays a pivotal role in the efflux of a wide variety of physiological substrates across the plasma membrane. Several studies have shown that MRP2 can significantly affect the absorption, distribution, metabolism, excretion, and toxicity (ADMET) profiles of many therapeutic drugs and chemicals found in the environment and diet. This transporter can also efflux newly developed anticancer agents that target specific signaling pathways and are major clinical markers associated with multidrug resistance (MDR) of several types of cancers. MDR remains a major limitation to the advancement of the combinatorial chemotherapy regimen in cancer treatment. In addition to anticancer agents, MRP2 reduces the efficacy of various drug classes such as antivirals, antimalarials, and antibiotics. The unique role of MRP2 and its contribution to MDR makes it essential to profile drug-transporter interactions for all new and promising drugs. Thus, this current research seeks to identify modulators of MRP2 protein expression levels using cell-based assays. A unique recently approved FDA library (372 drugs) was screened using a high-throughput In-Cell ELISA assay to determine the effect of these therapeutic agents on protein expression levels of MRP2. A total of 49 FDA drugs altered MRP2 protein expression levels by more than 50% representing 13.17% of the compounds screened. Among the identified hits, thirty-nine (39) drugs increased protein expression levels whereas 10 drugs lowered protein expression levels of MRP2 after drug treatment. Our findings from this initial drug screening showed that modulators of MRP2 peregrinate multiple drug families and signify the importance of profiling drug interactions with this transporter. Data from this study provides essential information to improve combinatorial drug therapy and precision medicine as well as reduce the drug toxicity of various cancer chemotherapies.
文摘目的探讨胆盐载体MRP1、MRP2在妊娠期肝内胆汁淤积症(ICP)胎盘的表达,分析胎盘胆盐载体与ICP发病的关系。方法收集8例正常早孕妇女绒毛组织、7例正常中孕妇女胎盘组织、20例正常晚孕妇女(对照组)胎盘组织,以及20例ICP患者胎盘组织,检测收集组织中胆盐载体MRP1和MRP2 mRNA的表达。结果ICP和正常妊娠各期胎盘组织中均有MRP2 mRNA的表达,而MRP1 mRNA在大部分标本上有表达;ICP组MRP1 mRNA和MRP2 mRNA表达量与对照组相比(99.94±73.17 vs 99.20±68.65;95.78±56.50 vs 142.20±91.27)差异无统计学意义(P>0.05);ICP组未用地塞米松治疗胎盘MRP2 mRNA表达量低于对照组(91.82±48.08 vs 142.20±91.27,P<0.05)。结论在未用地塞米松治疗的ICP患者胎盘组织中,MRP2 mRNA的表达量降低,这可能是引起ICP患者胎盘胆汁酸转运障碍,从而引起胎儿体内胆汁淤积的机制之一。
文摘根据猪链球菌2型(Streptococcus suis type 2,SS2)溶菌酶释放蛋白(muramidase-released protein,MRP)和胞外蛋白因子(extracellular protein factor,EPF)的基因序列,各设计合成一对引物,以猪链球菌2型江苏分离株SS2-1的基因组为模板,扩增mrp基因1 801~2 513位序列和epf 基因1 783~2 563位序列,分别构建原核表达载体pET32a-mrp 、pET32a-epf,确定诱导表达的两种蛋白都具有免疫原性后,提取阳性克隆质粒各自进行双酶切并纯化,通过PCR串联两片段,将目的片段定向克隆到表达载体pET-32a中,重组质粒转化入大肠杆菌BL21,经IPTG诱导表达分子量约为74kD的融合蛋白.用制备的两种抗血清与纯化融合蛋白进行免疫转印,结果显示融合蛋白分别具有MRP与EPF的中和表位.用融合蛋白MRP-EPF免疫新西兰兔,以最小致死量猪链球菌强毒株SS2-1攻击,兔的存活率达50%(2/4),存活率明显高于单个表达产物.证实串联表达的融合蛋白为重要的保护性抗原.
