TY - JOUR
T1 - Development of a quantitative diagnostic method of estrogen receptor expression levels by immunohistochemistry using organic fluorescent material-assembled nanoparticles
AU - Gonda, Kohsuke
AU - Miyashita, Minoru
AU - Watanabe, Mika
AU - Takahashi, Yayoi
AU - Goda, Hideki
AU - Okada, Hisatake
AU - Nakano, Yasushi
AU - Tada, Hiroshi
AU - Amari, Masakazu
AU - Ohuchi, Noriaki
N1 - Funding Information:
Part of this work was supported by a Grant-in-Aid for challenging Exploratory Research (21659144) by the Japan Society for the Promotion of Science (JSPS) (K. Gonda), Grant-in-Aid for Scientific Research (B) (24300336) by JSPS (K. Gonda), a Grant-in-Aid for Scientific Research in Innovative Areas “Nanomedicine Molecular Science” (23107009) from the Ministry of Education, Culture, Sports, Science, and Technology, Japan (K. Gonda), and a Grant-in-Aid for Comprehensive Research and Development of an Early Stage Diagnosis Method and Instruments to Treat Cancer from New Energy and Industrial Technology Development Organization, Japan (K. Gonda, M. Watanabe, Y. Nakano, N. Ohuchi). We also acknowledge the support of the Biomedical Research Core of Tohoku University Graduate School of Medicine and Konica Minolta Science and Technology Foundation.
PY - 2012/9/28
Y1 - 2012/9/28
N2 - The detection of estrogen receptors (ERs) by immunohistochemistry (IHC) using 3,3'-diaminobenzidine (DAB) is slightly weak as a prognostic marker, but it is essential to the application of endocrine therapy, such as 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.
AB - The detection of estrogen receptors (ERs) by immunohistochemistry (IHC) using 3,3'-diaminobenzidine (DAB) is slightly weak as a prognostic marker, but it is essential to the application of endocrine therapy, such as 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.
KW - Cancer
KW - Diagnosis
KW - Estrogen receptor
KW - Immunohistochemistry
KW - Nanoparticle
KW - Quantitative sensitivity
UR - http://www.scopus.com/inward/record.url?scp=84866888392&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84866888392&partnerID=8YFLogxK
U2 - 10.1016/j.bbrc.2012.08.105
DO - 10.1016/j.bbrc.2012.08.105
M3 - Article
C2 - 22959769
AN - SCOPUS:84866888392
VL - 426
SP - 409
EP - 414
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 3
ER -
