RRC ID 44189
Author Subramanian N, Raghunathan V, Kanwar JR, Kanwar RK, Elchuri SV, Khetan V, Krishnakumar S.
Title Target-specific delivery of doxorubicin to retinoblastoma using epithelial cell adhesion molecule aptamer.
Journal Mol. Vis.
Abstract PURPOSE:To study target-specific delivery of doxorubicin (Dox) using an RNA aptamer against epithelial cell adhesion molecule (EpCAM) in retinoblastoma (RB) cells.
METHODS:The binding affinity of the EpCAM aptamer to RB primary tumor cells, Y79 and WERI-Rb1 cells, and Müller glial cell lines were evaluated with flow cytometry. Formation of physical conjugates of aptamer and Dox was monitored with spectrofluorimetry. Cellular uptake of aptamer-Dox conjugates was monitored through fluorescent microscopy. Drug efficacy was monitored with cell proliferation assay.
RESULTS:The EpCAM aptamer (EpDT3) but not the scrambled aptamer (Scr-EpDT3) bound to RB tumor cells, the Y79 and WERI-Rb1 cells. However, the EpCAM aptamer and the scrambled aptamer did not bind to the noncancerous Müller glial cells. The chimeric EpCAM aptamer Dox conjugate (EpDT3-Dox) and the scrambled aptamer Dox conjugate (Scr-EpDT3-Dox) were synthesized and tested on the Y79, WERI-Rb1, and Müller glial cells. The targeted uptake of the EpDT3-Dox aptamer caused cytotoxicity in the Y79 and WERI-Rb1 cells but not in the Müller glial cells. There was no significant binding or consequent cytotoxicity by the Scr-EpDT3-Dox in either cell line. The EpCAM aptamer alone did not cause cytotoxicity in either cell line.
CONCLUSIONS:The results show that the EpCAM aptamer-Dox conjugate can selectively deliver the drug to the RB cells there by inhibiting cellular proliferation and not to the noncancerous Müller glial cells. As EpCAM is a cancer stem cell marker, this aptamer-based targeted drug delivery will prevent the undesired effects of non-specific drug activity and will kill cancer stem cells precisely in RB.
Volume 18
Pages 2783-95
Published 2012
PMID 23213278
PMC PMC3513190
MeSH Antibiotics, Antineoplastic / chemistry Antibiotics, Antineoplastic / pharmacology* Antigens, Neoplasm / genetics Antigens, Neoplasm / metabolism* Aptamers, Nucleotide / chemistry* Biological Transport Biomarkers, Tumor / genetics Biomarkers, Tumor / metabolism* Cell Adhesion Molecules / genetics Cell Adhesion Molecules / metabolism* Cell Line, Tumor Cell Proliferation / drug effects Cells, Cultured Doxorubicin / chemistry Doxorubicin / pharmacology* Drug Carriers / chemistry Drug Carriers / metabolism Drug Carriers / pharmacology* Epithelial Cell Adhesion Molecule Flow Cytometry Gene Expression Humans Molecular Targeted Therapy Neuroglia / cytology Neuroglia / drug effects Neuroglia / metabolism Organ Specificity Protein Binding Retinal Neoplasms / drug therapy Retinal Neoplasms / metabolism Retinal Neoplasms / pathology Retinoblastoma / drug therapy Retinoblastoma / metabolism Retinoblastoma / pathology Spectrometry, Fluorescence
IF 2.219
Times Cited 32
WOS Category OPHTHALMOLOGY BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Human and Animal Cells