Identification of somatostatin receptors using labeled PEGylated octreotide,as an active internalization

Abstract

Numerous normal and tumors cells are well-known to express the somatostatin receptors (SSTRs) on their surface which makes the receptor be useful for tumor scintigraphy. Thus, the identification of SSTRs is beneficial, especially SSTR₂. The somatostatin analog, Octreotide (OCT), was chosen as a ligand, as it is known to selectively bind to SSTR₂. Moreover, polyethylene glycol (PEG), 8armPEG, was used as a branched PEG to provide a low nonspecific cell binding and easily chemical modification. OCT and fluorescein (Flu) were conjugated to branched PEG using a water-soluble carbodiimide (EDC) and N-hydroxy succinimide (NHS) so as to activate its carboxylic acid group. 8armPEG-tagged Flu and OCT was characterized by gel permeation chromatography (GPC) to proof the conjugation of OCT to 8armPEG. Finally, cellular uptake was studied using pancreatic cancer cells with well-expressed somatostatin receptors using a confocal laser scanning microscope (CLMS) and fluorescence activated cell sorting (FACS). GPC showed increases in molecular mass since it showed a difference in elution time of 8armPEG itself and 8armPEG labeled with Flu. CLMS and FACS showed high binding with the positive SSTR₂ cells expression and showed negative results with negative expressing SSTR₂. These bindings were decreased when the receptors were occupied with free OCT which confirms the specific binding to SSTR₂. Therefore, we formulated a novel model to easily identify SSTR₂ and other receptors which serves as a promising platform for identification of tumor cells overexpressing the SSTR₂, which would be a hopeful target for cancer therapy and tumor scintigraphy.