Development of a quantitative diagnostic method of estrogen receptor expression levels by immunohistochemistry using organic fluorescent material-assembled nanoparticles
Development of a quantitative diagnostic method of estrogen receptor expression levels by immunohistochemistry using organic fluorescent material-assembled nanoparticles
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DOI：
10.1016/j.bbrc.2012.08.105
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发表时间：
2012-09-28
影响因子：
3.1
通讯作者：
Ohuchi, Noriaki
中科院分区：
文献类型：
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作者：
Gonda, Kohsuke;Miyashita, Minoru;Ohuchi, Noriaki
使用 3,3'-二氨基联苯胺 (DAB) 通过免疫组织化学 (IHC) 检测雌激素受体 (ER) 作为预后标志物稍弱，但它对于内分泌治疗的应用至关重要，例如基于芳香酶抑制剂或抗雌激素他莫昔芬的治疗。使用 DAB 的 IHC 是一种较差的定量方法，因为辣根过氧化物酶 (HRP) 活性取决于反应时间、温度和底物浓度。然而。使用荧光材料的 IHC 提供了定量使用 IHC 的有效方法，因为信号强度与光子激发能量的强度成正比。然而，高水平的自发荧光阻碍了利用荧光进行定量 IHC 的发展。我们开发了用于 IHC 的有机荧光材料（四甲基罗丹明）组装的纳米颗粒。组织自发荧光与量子点的荧光强度相当，量子点是最具代表性的荧光纳米颗粒。我们的新型纳米粒子的荧光强度比量子点高 10.2 倍，并且由于其表面覆盖有聚乙二醇链，因此它们不会非特异性地与乳腺癌组织结合。因此，我们的纳米颗粒的荧光强度显着超过自发荧光，这在 IHC 成像的癌症组织上产生了比以前的方法显着更高的信噪比。此外，我们在相邻组织切片的同一区域比较了纳米颗粒荧光 IHC 和 DAB IHC 的免疫染色数据，以定量检查这两种方法。结果表明，我们的纳米颗粒染色分析了广泛的 ER 表达水平，比 DAB 染色具有更高的准确性和定量灵敏度。使用我们的免疫染色方法提高 ER 诊断准确性和敏感性，将改善对下游 ER 信号诱导的针对 ER 和孕酮受体的治疗反应的预测。 (C) 2012 Elsevier Inc. 保留所有权利。
The detection of estrogen receptors (ERs) by immunohistochemistry (IHC) using 3,3'-diaminobenzidine (DAB) is slightly weak as a prognostic marker, but it is essentia to the application of endocrine therapy, such as aromatase inhibitor- or antiestrogen tamoxifen-based therapy. IHC using DAB is a poor quantitative method because horseradish peroxidase (HRP) activity depends on reaction time, temperature and substrate concentration. However. IHC using fluorescent material provides an effective method to quantitatively use IHC because the signal intensity is proportional to the intensity of the photon excitation energy. However, the high level of autofluorescence has impeded the development of quantitative IHC using fluorescence. We developed organic fluorescent material (tetramethylrhodamine)-assembled nanoparticles for IHC. Tissue autofluorescence is comparable to the fluorescence intensity of quantum dots, which are the most representative fluorescent nanoparticles. The fluorescent intensity of our novel nanoparticles was 10.2-fold greater than quantum dots, and they did not bind non-specifically to breast cancer tissues due to the polyethylene glycol chain that coated their surfaces. Therefore, the fluorescent intensity of our nanoparticles significantly exceeded autofluorescence, which produced a significantly higher signal-to-noise ratio on IHC-imaged cancer tissues than previous methods. Moreover, immunostaining data from our nanoparticle fluorescent IHC and IHC with DAB were compared in the same region of adjacent tissues sections to quantitatively examine the two methods. The results demonstrated that our nanoparticle staining analyzed a wide range of ER expression levels with higher accuracy and quantitative sensitivity than DAB staining. This enhancement in the diagnostic accuracy and sensitivity for ERs using our immunostaining method will improve the prediction of responses to therapies that target ERs and progesterone receptors that are induced by a downstream ER signal. (C) 2012 Elsevier Inc. All rights reserved.