RESUMO
The quantitative sensitivity and dynamic range of conventional immunohistochemistry (IHC) with 3,3'-diaminobenzidine (IHC-DAB) used in pathological diagnosis in hospitals are poor, because enzyme activity can affect the IHC-DAB chromogenic reaction. Although fluorescent IHC can effectively increase the quantitative sensitivity of conventional IHC, tissue autofluorescence interferes with the sensitivity. Here, we created new fluorescent nanoparticles called phosphor-integrated dots (PIDs). PIDs have 100-fold greater brightness and a more than 300-fold greater dynamic range than those of commercially available fluorescent nanoparticles, quantum dots, whose fluorescence intensity is comparable to tissue autofluorescence. Additionally, a newly developed image-processing method enabled the calculation of the PID particle number in the obtained image. To quantify the sensitivity of IHC using PIDs (IHC-PIDs), the IHC-PIDs method was compared with fluorescence-activated cell sorting (FACS), a method well suited for evaluating total protein amount, and the two values exhibited strong correlation (R = 0.94). We next applied IHC-PIDs to categorize the response to molecular target-based drug therapy in breast cancer patients. The results suggested that the PID particle number estimated by IHC-PIDs of breast cancer tissues obtained from biopsy before chemotherapy can provide a score for predicting the therapeutic effect of the human epidermal growth factor receptor 2-targeted drug trastuzumab.
Assuntos
Neoplasias da Mama/diagnóstico por imagem , Neoplasias da Mama/tratamento farmacológico , Diagnóstico por Imagem/métodos , Corantes Fluorescentes/química , Nanopartículas/química , Rodaminas/química , 3,3'-Diaminobenzidina/química , Anticorpos/química , Antineoplásicos Imunológicos/uso terapêutico , Biópsia , Biotina/química , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Diagnóstico por Imagem/instrumentação , Feminino , Fluorescência , Expressão Gênica , Humanos , Imidas/química , Imuno-Histoquímica/métodos , Pessoa de Meia-Idade , Proteínas de Neoplasias/antagonistas & inibidores , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Tamanho da Partícula , Perileno/análogos & derivados , Perileno/química , Receptor ErbB-2/antagonistas & inibidores , Receptor ErbB-2/genética , Receptor ErbB-2/metabolismo , Estreptavidina/química , Trastuzumab/uso terapêuticoRESUMO
In the past, dendritic polyphenylazomethines (DPA) have been synthesized via the convergent method. However, the convergent method has problems such as difficult terminal-group modifications and an increased number of steps. Therefore, we synthesized the terminal-protected DPA to construct the dendritic structure via the divergent method. The combination of the convergent method and the divergent method easily achieved the rapid synthesis of a higher generation dendrimer. Furthermore, we succeeded in the synthesis of the water-soluble PEG-DPA via the divergent method using the ester-DPA.
RESUMO
We have developed a novel synthetic method of phenylazomethine dendrons that uses 4,4'-methylenedianiline instead of 4,4'-diaminobenzophenone to synthesize the precursor of the phenylazomethine dendron and then oxidized the precursor to the next-generation dendron. For this method, the productivity of the dendrons has been significantly increased. Furthermore, as the synthesis of high-generation dendrons becomes easier, synthesis of DPA G5 was achieved. [structure--see text]
