Evaluation of ssDNA aptamer binding to Aurora Kinase A and its cytotoxic effects on HeLa and MCF-7 cancer cell lines

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Purpose: To investigate the binding potential of ssDNA aptamer to Aurora kinase A (AURKA) and assess its inhibitory effect on HeLa and MCF-7 cell lines. Methods: A detailed analysis was conducted on the binding between aptamer and AURKA protein using HDOCK and HADDOCK, with 2D interaction obtained by LIGPLOT+ v1.4. Binding of FAM-labelled ssDNA aptamer to AURKA was analyzed by EVOS-5000 imaging and the inhibitory effects of aptamer and aptamer-drug complexes (Bleomycin Sulfate, Metformin, and Mitomycin C) at different concentrations and times were evaluated by PrestoBlue assay. Results: Docking results showed a HADDOCK score of -82.0 ± 6.4 and a Z-score of -1.5. The HDOCK result showed best docking result (-327.13) at a confidence interval of 0.9719. Free aptamer and its drug complexes effectively inhibited HeLa cell growth, with the highest cytotoxic effect observed at higher concentrations. Free aptamer achieved the highest drug sensitivity score (DSS: 40.7) in HeLa, followed by its bleomycin sulfate and metformin complex (DSS: 31.6). However, MCF-7 exhibited lower sensitivity and DSS score. Conclusion: ssDNA aptamer shows potential in cancer detection and therapy. In vitro and in vivo studies are needed to ascertain the potential of ssDNA aptamers in cancer therapy.

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