Evaluation of the Cytotoxicity of a Circular SIRT1 Aptamer on MCF-7 and A549 Cell Lines and Its Impact on HIF-1α and VEGFR2 Expression
DOI:
https://doi.org/10.32947/ajps.v26i3.1407Keywords:
SIRT1 aptamer, A549, MCF-7, HUVECs, cell viability, relative selectivity index, XTT assayAbstract
Background: Selective cytotoxicity against malignant cells with limited toxicity to normal cells remains an important goal in anticancer drug development.
Objectives: To evaluate the cell viability and preliminary relative selectivity of a circular single stranded DNA SIRT1 aptamer in A549 human lung adenocarcinoma cells and MCF-7 human breast adenocarcinoma cells compared with HUVECs human umbilical vein endothelial cells, and to study its effects on HIF-1α and VEGFR2 protein expression.
Methods: A549 and MCF-7 cells were treated with SIRT1 aptamer concentrations ranging from 0.0195 to 10 µM for 24, 48, and 72 h, while HUVECs were treated with concentrations ranging from 0.078 to 40 µM for the same exposure times. Cell viability was measured by XTT assay. IC50 and selectivity index values using HUVECs as the non-malignant comparator were calculated. HIF-1α and VEGFR2 expression in A549 and MCF-7 cells were measured by Western blotting.
Results: The aptamer induced time- and concentration-dependent reduction in cell viability. The IC50 values after 72 h were 0.742 µM in A549, 0.473 µM in MCF-7 and 4.018 µM in HUVECs. The selectivity index values were 5.42 and 8.49 for A549 and MCF-7 respectively. HIF-1α was significantly reduced in both cancer cell lines, while VEGFR2 was significantly reduced only in A549 cells.
Conclusion: The circular SIRT1 aptamer showed preliminary relative selectivity against A549 and MCF-7 cancer cell lines compared with HUVECs and was associated with reduced HIF-1α expression, with VEGFR2 reduction observed mainly in A549 cells. However, further studies are required to confirm SIRT1 involvement, define the mechanism of cell-death/viability reduction, and determine whether these molecular changes translate into functional antiangiogenic effects.
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