Somatostatin receptor staining in FFPE sections using a ligand derivative dye as an alternative to immunostaining.
The confirmation of target expression in tissues is a prerequisite for molecular-targeted therapy. However, difficulties are sometimes associated with the production of appropriate antibodies against receptors. We herein developed a ligand derivative dye for the staining of receptors. The somatostatin receptor (sstr) was selected as the target and FITC-octreotate as the detective agent. We performed a blot analysis to detect sstr in the transfer membrane. The sstr2 recombinant protein or cell lysate from a small cell lung carcinoma cell line (H69) was boiled and loaded onto SDS-PAGE, and the proteins were transferred to a membrane. Even after denaturing processes, FITC-octreotate still bound sstr on the membrane. Furthermore, FITC-octreotate depicted the expression of sstr in formalin-fixed and paraffin-embedded (FFPE) sections, a method that we named ligand derivative staining (LDS). The accuracies of immunostaining and LDS were compared at the points of the detection of sstr using FFPE sections of 30 neuroendocrine tumor specimens. The sensitivity of LDS was 81.8%, while those of immunostaining using anti-sstr2 and sstr5 antibodies were 72.7% and 63.6%, respectively. Thus, LDS appears to be superior to immunostaining. A ligand derivative may be used as a substitute for antibodies, and has the potential to support economical, simple, and accurate detection methods.
Department of Pathology and Experimental Medicine, Kumamoto University, Graduate School of Medical Sciences, Kumamoto, Japan.|Center for Instrumental Analysis, Kyoto Pharmaceutical University, Kyoto, Japan.
Koki
http://orcid.org/0000-0002-1479-2534
K
Hasegawa
Department of Pathology and Experimental Medicine, Kumamoto University, Graduate School of Medical Sciences, Kumamoto, Japan.
Shinji
S
Kudoh
Department of Pathology and Experimental Medicine, Kumamoto University, Graduate School of Medical Sciences, Kumamoto, Japan.
Takaaki
T
Ito
D045744:Cell Line, Tumor; D016650:Fluorescein-5-isothiocyanate / Q000494:pharmacology; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D018358:Neuroendocrine Tumors / Q000378:metabolism* / Q000473:pathology; D015282:Octreotide / Q000031:analogs & derivatives* / Q000494:pharmacology; D011485:Protein Binding; D017481:Receptors, Somatostatin / Q000378:metabolism*; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods*; D016610:Tissue Embedding / Q000379:methods
D017481:Receptors, Somatostatin; D016650:Fluorescein-5-isothiocyanate; D015282:Octreotide