Abstract
This study explores the synergistic effect of T-CUS245C combined with cyclosporin A, and its possible mechanism. T-CUS245C is an immunotoxin conjugated with tastuzumab and recombinant cucurmosin 245C (CUS245C). Cyclosporin A is a calcium antagonist mainly used as an immune-suppressive agent, which enhances the cytotoxicity of immunotoxins through an unknown mechanism. The sulforhodamine B assay, flow cytometry, DNA fragmentation apoptosis assay, and western blot analysis were used to evaluate the synergistic effects of cyclosporin A combined with T-CUS245C on proliferation inhibition and apoptosis induction in SK-OV-3 ovarian cancer cells with HER-2-overexpressing. Meanwhile, using confocal microscopy and flow cytometry, the effect of cyclosporin A on the intracellular distribution of T-CUS245C was observed and quantified. The results of this study showed that the combination index of cyclosporin A combined with T-CUS245C is less than 0.7, indicating that cyclosporin A significantly enhances the proliferation inhibition and apoptosis induction induced by T-CUS245C in SK-OV-3 cells. The confocal microscopy and flow cytometry results show that cyclosporin A effectively increases the dispersion of T-CUS245C in the cytoplasm by promoting the endolysosomal escape. This study demonstrates that cyclosporin A combined with T-CUS245C has a synergistic anticancer effect, by facilitating the release of T-CUS245C in the intracellular space.
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The data that support the findings of this study are available from the corresponding author upon reasonable request.
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Acknowledgements
We thank the public technology service center of Fujian Medical University for helping with the experiment.
Funding
This research was funded by the National Science Foundation of China (No. 30772587), Science Foundation of State Key Laboratory of Structural Chemistry (No. 20210001), Fujian Provincial Central Leading Local Science and Technology Development Fund Project (No. 2021L3025), the Natural Science Foundation of Fujian Province (No. 2020J01436), the Fujian Province Health and Family Planning Scientific Research Talent Training Project (No. 2018-CX-40), Young and Middle-aged Scientific Research Project of Ningde Normal University (No. 2020Q103), the Startup Fund for Scientific Research (No. 2018QH1013), and the Innovation Team project of Ningde Normal University (No. 2022T03).
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Study concept and design: HX, JX, XY; acquisition of data: HX, YF; analysis and interpretation of data: HX, ZL, YF; drafting of the manuscript: HX; critical revision of the manuscript for important intellectual content: JX, XY; statistical analysis: HX, YF; obtained funding: JX, XY; administrative, technical, or material support: JX; study supervision: JX, XY.
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Xie, H., Fu, Y., Lin, Z. et al. Cyclosporin A Enhance the Cytotoxicity of Immunotoxin to the HER-2-Overexpressing SK-OV-3 Cells: A Prospective Study. Rev. Bras. Farmacogn. (2024). https://doi.org/10.1007/s43450-024-00551-6
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DOI: https://doi.org/10.1007/s43450-024-00551-6