Investigation of the therapeutic effi̇cacy of chitosansilver nanoparticle-loaded doxorubicin on liver cancer cells

Nanobiotechnology is seen as a reliable approach to overcome most of the problems related to anticancer drug delivery. The development of therapeutics and therapeutic nano-drug delivery systems is a necessity to have a variety of nanocarriers and active substances for better treatment and efficient controlled release of drugs on a single platform and is the research topic of the current project proposal. Therapeutics and therapeutic systems have advantages over conventional chemotherapy due to high efficiency of drug loading, high cellular uptake, high drug release and minimal side effects. The use of chitosan-silver nanoparticles (K-AgNPs) in the treatment of various cancer types is being investigated due to their superior properties. This study aims to investigate the cytotoxic, proliferative and apoptotic effects of K-AgNP loaded Doxorubicin (Dox) on liver cancer cells (HepG2). In the study, after the synthesis and characterisation of K-AgNP, Dox loading was provided and the release of the loading determined by FITR, UV-Vis, XRD, SEM methods was analysed. Then, cell viability analysis of the obtained K-AgNP+Dox was performed and inhibitory concentration was determined. Then, two therapeutic doses (low and high) under the IC50 value were determined and the applications were started. Incubation periods of 24 and 48 hours were provided for all applications and various gene and protein expressions were determined from the cells obtained by RT-PCR and western blot method. In the analysis of the data obtained, it was determined that the IC50 value of K-AgNP+Dox was 50 µg/mL for 24h and 30 µg/mL for 48h. AKT, PI3K, EGFR, BAX and Bcl2 gene expressions were determined in the obtained RNA samples. In Western blot analysis, the expression levels of NFKB-p65, LC3, Beclin1, Bcl-2 and Caspase-3 proteins were determined. The findings of our study revealed that administration of K-AgNP-loaded Dox significantly inhibited the proliferation of liver cancer cells (HEPG2) and enhanced their apoptotic response. This suggests that K-AgNP-mediated delivery of Dox effectively improves its therapeutic efficacy, potentially by enhancing targeted delivery and cellular uptake of the drug.