Objective: To explore the effects of siRNA-mediated downregulation of integrin-linked kinase (ILK) gene expression on the proliferation and apoptosis in retinoblastoma cells.
Methods: Experiment study. Human retinoblastoma cells, HXO-Rb(44) cells, were divided into four groups: ILK siRNA intervention group, control siRNA intervention group, empty liposome group and blank control group. ILK siRNA was transfected into HXO-Rb(44) cells by lipofection. The expression of ILK mRNA and protein was detected at 48 hours after transfection by RT-PCR and Western blot, respectively. Cell proliferation inhibition was measured by Cell Counting Kit-8 assay. The apoptosis was detected by Annexin/PI double immunofluorescence and flow cytometry. Statistical method adopted one-way ANOVA between each group overall comparison.
Results: The HXO-Rb(44) cells were transfected by ILK siRNA successfully with lipofection. RT-PCR analysis showed that the expression of ILK mRNA in the ILK siRNA intervention group was significantly decreased as compared to the control siRNA intervention group, empty liposome group and blank control group (0.12 ± 0.02 vs. 0.45 ± 0.04, 0.42 ± 0.05 and 0.40 ± 0.04, F = 12.781, P = 0.000). Similar results were obtained for protein expression as revealed by Western blot analysis (0.10 ± 0.03 vs. 0.38 ± 0.07, 0.40 ± 0.05 and 0.43 ± 0.03, F = 18.647, P = 0.000). The proliferation inhibition rate in the ILK siRNA intervention group (35.0%) was higher than that in the blank control group (0%), control siRNA intervention group (2.1%) and empty liposome group (1.8%) (F = 23.573, P = 0.000). The results of immunofluorescence and flow cytometry showed that ratio of Annexin positive cells was highest in the ILK siRNA intervention group (26.5%), which was 5.0%, 4.6%, 5.3% in the empty liposome group, blank control group, control siRNA intervention group, respectively (F = 65.217, P = 0.000).
Conclusion: ILK siRNA can downregulate the expression of ILK gene in HXO-Rb(44) cells inhibited the cell proliferation and induced their apoptosis.