Abstract
P2Et extract obtained from the Caesalpinia spinosa plant is abundant in phenolic compounds such as gallic acid and ethyl gallate and can generate signals to activate the immune response by inducing a mechanism known as immunogenic cell death in murine models of breast cancer and melanoma. Immunogenic cell death involves mechanisms such as autophagy, which can be modulated by various natural compounds, including phenolic compounds with a structure similar to those found in P2Et extract. Here, we determine the role of autophagy in apoptosis and the generation of immunogenic signals using murine wild-type B16-F10 melanoma cells and cells with beclin-1 gene knockout. We show that P2Et extract and ethyl gallate induced autophagy, partially protecting tumor cells from death and promoting calreticulin exposure and the release of ATP. Although ethyl gallate showed a mechanism similar to that of P2Et, the induction of apoptosis and immunogenic signals was significantly weaker. In contrast, gallic acid-induced autophagy acted by blocking autophagic flux, which was associated with increased cell death. However, this compound did not induce any of the immunogenic death signals evaluated. Therefore, the complex extract has greater antitumor potential than isolated compounds. Here, we show that inducing autophagic flux with P2Et protects cancer cells from cell death and that this delay in cell death is required for the generation of immunogenic signals.
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Acknowledgments
This project has the authorization of the Ministry of the environment, for the use of genetic resources through the Contract of Access to Genetic Resources No. 220/2018 for Colombian plant material. The authors want to express their gratitude to Paola Lasso for the edition of the figures.
Funding
Funding was provided by the Departamento Administrativo de Ciencia, Tecnología e Innovación COLCIENCIAS (120356934797, contract number 701–2013) and Vicerrectoría de Investigaciones, Pontificia Universidad Javeriana (007682 and 009067) Bogotá, Colombia. K.P was funded by the Departamento Administrativo de Ciencia, Tecnología e Innovación COLCIENCIAS, convocatoria 647 de 2014.
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KP carried out the experiments, analyzed data and co-wrote the paper. MPL carried out the lysosomal damage experiments. CU was involved in planning, analyzed data and supervised the work. CJA-D was involved in performed, planning and supervised the generation of knockout cell lines. AB and SF conceived the original idea supervised the project and prepared the manuscript. All authors discussed the results and contributed to the final manuscript.
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SF and CU are inventors of a granted patent related to P2Et. The remaining authors declare that the research was conducted in the absence of any potential conflict of interest.
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Prieto, K., Lozano, M.P., Urueña, C. et al. The delay in cell death caused by the induction of autophagy by P2Et extract is essential for the generation of immunogenic signals in melanoma cells. Apoptosis 25, 875–888 (2020). https://doi.org/10.1007/s10495-020-01643-z
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DOI: https://doi.org/10.1007/s10495-020-01643-z