Investigation of the combined therapeutic use of rna-based P53 tumor suppressor gene activation and kras proto-oncogene inhibition in the A549 human lung adenocarcinoma cell line

Aim: The study aimed to reveal the anticancer effect of the combined use of siRNA, which inhibits KRAS expression, and saRNA, which activates p53 expression, in cancer cells carrying mutant KRAS and wild-type p53, compared to their separate transfections, and to develop a new RNA-based therapeutic approach. Materials and Methods: In the study, A549 cell line was used as the basis and HCT116 cell line was used as a control. AuNP synthesis was performed, the synthesized AuNPs were conjugated with small RNAs and the cells were transfected with the conjugates. Post-transfection mRNA and protein expression analyzes were performed by RT-qPCR and Western Blot methods, respectively. Effective small RNA concentrations were determined by mRNA and protein analyses. Combined and separate transfection processes were carried out using the determined concentrations, and the events at the cellular level caused by transfection were examined by performing cell viability test, cell cycle analysis, apoptosis and necrosis test, colony formation experiment, wound healing experiment, invasion and migration experiments. The results were evaluated statistically by applying Student's t-test. Results: Co-transfection of small RNAs at a concentration of 50 nM was found to result in a significant increase in p53 mRNA levels (p<0.001, p<0.01) and protein levels (p<0.001, p<0.01), and did not alter KRAS mRNA and protein levels (p>0.05) in the A549 and HCT116 cell lines, respectively, compared to their individual transfections. In both cell lines, it was determined that the combined treatment approach more effectively arrested the cell cycle at the G0/G1 phase, triggered apoptosis to a greater extent, and significantly reduced cell proliferation, colony formation ability, invasion, and migration compared to individual transfections. Conclusion: It has been demonstrated that the combined approach subject to our study provides an anticancer effect for both cell lines compared to the control group, and that this approach has a much greater anticancer effect than the anticancer effect provided by both small RNAs separately. Key Words: A549, HCT116, siRNA, saRNA, KRAS, p53.