Tens of thousands of protein-protein interactions (PPIs) have been found in human cells and many of these macromolecular partnerships could determine the cell growth and death. Thus there is a need to develop the meth...Tens of thousands of protein-protein interactions (PPIs) have been found in human cells and many of these macromolecular partnerships could determine the cell growth and death. Thus there is a need to develop the methods to catalogue these macromolecules by detecting their interactions, modifications, and cellular locations. It will be helpful for scientists to compare the difference between a diseased cellular state and its normal state and to find the potential therapy treatment to intervene this status. One technology called split-protein reassembly or protein fragment complementation has been developed in the last two decades. This technology makes use of appropriate fragmentation of some protein reporters and the refolding of these reports could be detected by their function to confirm the interaction of interest. This system has been set up in cell-free systems, </span><i><span style="font-family:Verdana;">E.</span></i></span><i><span style="font-family:""> </span></i><i><span style="font-family:Verdana;">coli</span></i><span style="font-family:""><span style="font-family:Verdana;">, yeast, mammalian cells, plants and live animals. Herein, I present the development in fluorescence- and bioluminescence-based split-protein biosensors in both binary and ternary systems. In addition, some people developed the split-protein system by combining it with chemical inducer of dimerization strategy (CID). This has been applied for identifying the enzyme inhibitors and regulating the activity of protein kinases and phosphatases. With effort from many laboratories from the world, a variety of split-protein systems have been developed for studying the PPI </span><i><span style="font-family:Verdana;">in vitro</span></i><span style="font-family:Verdana;"> and </span><i><span style="font-family:Verdana;">in vivo</span></i><span style="font-family:Verdana;">, monitoring the biological process, and controlling the activity of the enzyme of interest.展开更多
目的探讨分裂相关增强子1(Hairy and enhancer of split related protein 1,HESR-1)、细胞分裂周期蛋白25同源蛋白C(Cell division cycle 25C,CDC25C)在直肠癌组织中的表达及其临床意义。方法选择166例行根治性手术的直肠癌患者为研究对...目的探讨分裂相关增强子1(Hairy and enhancer of split related protein 1,HESR-1)、细胞分裂周期蛋白25同源蛋白C(Cell division cycle 25C,CDC25C)在直肠癌组织中的表达及其临床意义。方法选择166例行根治性手术的直肠癌患者为研究对象,采用免疫组织化学法检测癌组织及癌旁组织中HESR-1和CDC25C的表达,收集患者临床病理参数并进行随访,分析HESR-1和CDC25C表达对直肠癌患者临床病理参数及预后的影响。结果肠癌组织中CDC25C、HESR-1的阳性率均高于癌旁组织(46.9%vs 12.6%,41.6%vs 7.6%),差异有统计学意义(P<0.05)。在癌组织中,CDC25C mRNA与HESR-1 mRNA的表达呈正相关(r=0.862,P=0.003)。癌组织CDC25C及HESR-1表达阳性患者的肿瘤直径和淋巴结转移率均高于表达阴性者(P<0.05)。Cox多因素分析结果显示,肿瘤直径、淋巴结转移、CDC25C阳性及HESR-1阳性是影响直肠癌患者预后的独立危险因素(P<0.05)。结论HESR-1和CDC25C在直肠癌组织中表达升高,且与患者预后密切相关。展开更多
文摘Tens of thousands of protein-protein interactions (PPIs) have been found in human cells and many of these macromolecular partnerships could determine the cell growth and death. Thus there is a need to develop the methods to catalogue these macromolecules by detecting their interactions, modifications, and cellular locations. It will be helpful for scientists to compare the difference between a diseased cellular state and its normal state and to find the potential therapy treatment to intervene this status. One technology called split-protein reassembly or protein fragment complementation has been developed in the last two decades. This technology makes use of appropriate fragmentation of some protein reporters and the refolding of these reports could be detected by their function to confirm the interaction of interest. This system has been set up in cell-free systems, </span><i><span style="font-family:Verdana;">E.</span></i></span><i><span style="font-family:""> </span></i><i><span style="font-family:Verdana;">coli</span></i><span style="font-family:""><span style="font-family:Verdana;">, yeast, mammalian cells, plants and live animals. Herein, I present the development in fluorescence- and bioluminescence-based split-protein biosensors in both binary and ternary systems. In addition, some people developed the split-protein system by combining it with chemical inducer of dimerization strategy (CID). This has been applied for identifying the enzyme inhibitors and regulating the activity of protein kinases and phosphatases. With effort from many laboratories from the world, a variety of split-protein systems have been developed for studying the PPI </span><i><span style="font-family:Verdana;">in vitro</span></i><span style="font-family:Verdana;"> and </span><i><span style="font-family:Verdana;">in vivo</span></i><span style="font-family:Verdana;">, monitoring the biological process, and controlling the activity of the enzyme of interest.
文摘目的探讨分裂相关增强子1(Hairy and enhancer of split related protein 1,HESR-1)、细胞分裂周期蛋白25同源蛋白C(Cell division cycle 25C,CDC25C)在直肠癌组织中的表达及其临床意义。方法选择166例行根治性手术的直肠癌患者为研究对象,采用免疫组织化学法检测癌组织及癌旁组织中HESR-1和CDC25C的表达,收集患者临床病理参数并进行随访,分析HESR-1和CDC25C表达对直肠癌患者临床病理参数及预后的影响。结果肠癌组织中CDC25C、HESR-1的阳性率均高于癌旁组织(46.9%vs 12.6%,41.6%vs 7.6%),差异有统计学意义(P<0.05)。在癌组织中,CDC25C mRNA与HESR-1 mRNA的表达呈正相关(r=0.862,P=0.003)。癌组织CDC25C及HESR-1表达阳性患者的肿瘤直径和淋巴结转移率均高于表达阴性者(P<0.05)。Cox多因素分析结果显示,肿瘤直径、淋巴结转移、CDC25C阳性及HESR-1阳性是影响直肠癌患者预后的独立危险因素(P<0.05)。结论HESR-1和CDC25C在直肠癌组织中表达升高,且与患者预后密切相关。