SiRNA-mediated Silencing of the RPS19 Gene Induces Apoptosis and Inhibits Cell Cycle Progression in Chronic Myeloid Leukemia Cells

Abstract

This research delves into the therapeutic implications of utilizing small interfering RNA (siRNA) to target the ribosomal protein S19 (RPS19) gene in chronic myeloid leukemia (CML) using the K562 cell line model. The primary objective was to investigate how gene silencing affects apoptosis promotion and cell cycle arrest. The study employed bioinformatics tools and databases to explore the interactions involving RPS19 and neighboring proteins. Subsequently, siRNA-mediated gene silencing was utilized to suppress RPS19 expression in K-562 cells, with assessments conducted on cell cycle progression and apoptosis through flow cytometry analysis. Furthermore, real-time PCR was employed to evaluate the expression levels of RPS19, along with the closely associated RPS16 and RPS18 genes. Silencing the RPS19 gene in siRNA-transfected K-562 cells led to an increase in apoptotic cells by over 20%, with a significant accumulation in the sub-G1 and G1 phases. Additionally, the knockdown of RPS19 resulted in a 75% decrease in RPS16 expression and a 50% decrease in RPS18 expression. These results demonstrate the therapeutic potential of targeting RPS19 in CML cells, suggesting a promising approach for precise treatment strategies in leukemia and potentially other types of cancer.

Keywords: RPS19; apoptosis; cell cycle; chronic myeloid leukemia; gene silencing; siRNA.

Fig. 1

The details of the interaction of RPS19 protein with RPS16 and RPS18 proteins. a) Spatial arrangement of RPS18 (purple) and RPS16 (green) proteins on both sides of RPS19 protein. Normal RPS19 protein is marked in blue and mutant protein is marked in gray. b) Residues involved in the RPS19 and RPS18 interaction are shown in yellow. c) Residues involved in the RPS19 and RPS16 interaction are shown in red. (source: http://www.uniprot.org).

Fig. 2

The expression level of RPS16 (a), RPS18 (b), and PRS19 (c) genes in K-562 cells transfected with RPS19-specific siRNA after 48 hours of incubation compared to normal cells. (*P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001).

Fig. 3

The effect of siRNA targeting RPS19 on the apoptosis rate of K-562 leukemia cells after 48 hours of incubation (b) compared to the nontreated cells (a). Q1: Necrosis, Q2: Late Apoptosis, Q3: Early Apoptosis, Q4: Live Cells.

Fig. 4

The effect of RPS19 knockdown on the cell phase of K-562 leukemia cells after 48 hours. a) Histogram of the control group, b) siRNA-treated K-562 cells group, and c) bar plot analysis of the cell phase of K-562 cells treated with siRNA against RPS19 compared to control cells. The analysis was calculated by the use of the student t-test. P-value < 0.05 was displayed as statistical significance. (*P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001).