Development and characterization of gene material loaded nanocarrier systems for lung cancer treatment

Cancer is a genetic defect disease. Gene therapy is one of the targeted treatment approaches in cancer and has the potential to eliminate the genetic cause of the disease. It has been discovered that lncRNA-UCA1, a type of non-coding RNA, is highly expressed in lung cancer. The aim of this thesis is to develop and characterize chitosan nanoparticles, which, thanks to the antisense oligonucleotides (GapmeRs) they contain, reduce the level of lncRNA-UCA1 and thus may be effective in lung cancer. Nanoparticles were prepared by ionic gelation method by combining chitosan and tripolyphosphate. Unloaded nanoparticles were optimized in terms of particle size and zeta potential by changing some formulation parameters. The optimum storage temperature for unloaded nanoparticle dispersions was found to be 5°C ± 3°C. As a result of optimization and dispersion stability studies, the selected values of the formulation parameters were chitosan/tripolyphosphate mass ratio [5.0:1], chitosan solution pH 4.0 and stirring speed 600 rpm. Particle size and zeta potential values were 164.90±2.51 nm and 39.60±0.81 mV for unloaded nanoparticles; 149.00±0.36 nm and +32.20±0.87 mV for GapmeR loaded nanoparticles. The encapsulation efficiency is 99.42 ± 1.10%. Agarose gel electrophoresis showed that chitosan nanoparticles and GapmeRs have good affinity. It has been shown that the nanoparticles are not cytotoxic to Calu-3 cells at the applied concentration. In the confocal microscopy study, it was observed that GapmeRs were taken into the cells. GapmeR-loaded nanoparticle application resulted in a significant decrease in lncRNA-UCA1 level only after 72 hours compared to the control groups.