Akademik Veri Yönetim Sistemi
9th International Congress of Molecular Biology Association of Turkey, İzmir, Türkiye, 14 Eylül 2024, ss.83
Background/Aim: Small RNAs are used in many studies, including the investigation of cancer treatments,
to modulate gene expression both in terms of activation and inhibition. For our study that
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 human lung adenocarcinoma cell line was used as a
model. The cells were transfected with small RNAs by using gold nanoparticules. Post-transfection
mRNA and protein expression analyzes were performed by RT-qPCR and Western Blot methods,
respectively. 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) and protein levels (p<0.001), and did not alter KRAS
mRNA and protein levels (p>0.05) in the A549 cell line, compared to their individual transfections.
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 human lung cancer cell line compared to the control group, and that this approach
has a much greater anticancer effect than the anticancer effect provided by each of the small RNAs
separately.
Keywords: p53, saRNA, KRAS, siRNA, A549