Inhibition of EGFR expression with chitosan/alginate nanoparticles encapsulating antisense oligonucleotides in T47D cell line using RT-PCR and immunocytochemistry

Chitosan/alginate nanoparticles optimized for size and loading efficiency were evaluated for their potential of antisense oligonucleotide delivery. The antisense for epidermal growth factor receptor (EGFR) that is over-expressed in many cancer cells was loaded in chitosan/alginate nanoparticles. The T47D breast cancer cell line was chosen to study the efficiency of optimized nanoparticles (chitosan/alginate 1:1, alginate/calcium chloride 0.2% and N/P ratio of 5 and 25) on EGFR expression. The MTT cytotoxicity evaluation of nanoparticles confirmed non-toxic properties of these carriers. The FITC-labeled EGFR antisense showed that T47D cells can uptake antisense-loaded nanoparticles better than naked antisense. Both RT-PCR and immunocytochemistry analyses showed that nanoparticles with N/P ratio of 5 can downregulate the expression of EGFR in T47D breast cancer cell line by improving internalization and stability of antisense molecules.