EpCAM aptamer mediated cancer cell specific delivery of EpCAM siRNA using polymeric nanocomplex
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RESEARCH
Open Access
EpCAM aptamer mediated cancer cell specific delivery of EpCAM siRNA using polymeric nanocomplex Nithya Subramanian1,2, Jagat R Kanwar2*, Prasanna kumar Athalya1,2, Narayanan Janakiraman1, Vikas Khetan4, Rupinder K Kanwar2, Sailaja Eluchuri1 and Subramanian Krishnakumar1,3*
Abstract Background: Epithelial cell adhesion molecule (EpCAM) is overexpressed in solid tumors and regarded as a putative cancer stem cell marker. Here, we report that employing EpCAM aptamer (EpApt) and EpCAM siRNA (SiEp) dual approach, for the targeted delivery of siRNA to EpCAM positive cancer cells, efficiently inhibits cancer cell proliferation. Results: Targeted delivery of siRNA using polyethyleneimine is one of the efficient methods for gene delivery, and thus, we developed a novel aptamer-PEI-siRNA nanocomplex for EpCAM targeting. PEI nanocomplex synthesized with EpCAM aptamer (EpApt) and EpCAM siRNA (SiEp) showed 198 nm diameter sized particles by dynamic light scattering, spherical shaped particles, of 151 ± 11 nm size by TEM. The surface charge of the nanoparticles was −30.0 mV using zeta potential measurements. Gel retardation assay confirmed the PEI-EpApt-SiEp nanoparticles formation. The difference in size observed by DLS and TEM could be due to coating of aptamer and siRNA on PEI nanocore. Flow cytometry analysis revealed that PEI-EpApt-SiEp has superior binding to cancer cells compared to EpApt or scramble aptamer (ScrApt) or PEI-ScrApt-SiEp. PEI-EpApt-SiEp downregulated EpCAM and inhibited selectively the cell proliferation of MCF-7 and WERI-Rb1 cells. Conclusions: The PEI nanocomplex fabricated with EpApt and siEp was able to target EpCAM tumor cells, deliver the siRNA and silence the target gene. This nanocomplex exhibited decreased cell proliferation than the scrambled aptamer loaded nanocomplex in the EpCAM expressing cancer cells and may have potential for EpCAM targeting in vivo. Keywords: EpCAM, Aptamer, PEI-EpApt-SiEp, siRNA delivery, Cancer targeting
Background Epithelial cell adhesion molecule (EpCAM) is highly expressed in most of the solid tumors. It has been reported as a putative cancer stem cell marker [1,2], and is regarded as a target antigen for cancer therapies using antibody, ankyrins and aptamers [3,4]. Since an aptamer based tumor targeting can rescue the inherent issues * Correspondence: [email protected]; [email protected] 2 Nanomedicine Laboratory of Immunology and Molecular Biomedical Research (LIMBR), School of Medicine (SoM), Molecular and Medical Research (MMR) Strategic Research Centre, Faculty of Health, Deakin University, Geelong, Victoria 3217, Australia 1 Department of Nanobiotechnology, Vision Research Foundation, Kamalnayan Bajaj Institute for Research in Vision and Ophthalmology, 18 College Road, Chennai 600006, Tamil Nadu, India Full list of author information is available at the end of the article
associated with antibody, such as larger size, immunogenicity, both DNA and RNA aptamers against EpCAM were developed using Systemic Evolution of Li
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