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Dual Akt and Bcl-2 inhibition induces cell-type specific modulation of apoptotic and autophagic signaling in castration resistant prostate cancer cell lines

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Abstract

Background

Cancer cell survival depends on the cross-regulation between apoptosis and autophagy which share common signaling pathways including PI3K/Akt/mTOR and Bcl-2. The aim of this study was to elucidate the modulation patterns between apoptosis and autophagy following dual inhibition by Akt inhibitor erufosine and Bcl-2 inhibitor ABT-737 in castration-resistant prostate cancer (CRPC) cell lines, PC-3 (Bax+) and DU-145 (Bax−).

Methods and Results

Cell cycle progression, apoptotic and autophagic signaling were examined by flow cytometry, multi-caspase assay, Hoechst staining, acridine orange staining of acidic vesicular organelles (AVOs), qRT-PCR and Western Blot. Dual inhibition increased G2/M arrest in PC-3 and DU-145, but not in the healthy prostate epithelium cells, PNT-1A. Only in PC-3, dual inhibition induced synergistic apoptotic and additive autophagic effects. In DU-145 and PNT-1A cells, ABT-737 did not display any remarkable effect on multicaspase activity and erufosine and ABT-737, neither alone nor in combination induced AVOs. By dual inhibition, AKT, BCL-2 and NF-κB gene expressions were downregulated in PC-3, both ATG-5 and BECLIN-1 gene expressions were upregulated in DU-145 but Beclin-1 protein expression was substantially reduced in both CRPC cells. Dual inhibition-induced synergistic multicaspase activation in PC-3 degrades and disrupts autophagic activity of Beclin-1, enhancing caspase-dependent apoptosis. However, in DU-145, following dual inhibition, rate of multicaspase induction and apoptosis are lower but autophagy is completely abolished despite markedly increased BECLIN-1 gene expression.

Conclusion

In conclusion, antineoplastic drug combinations may display cell-type specific modulation of apoptotic and autophagic signaling and lack of protective autophagy may not necessarily indicate increased chemotherapeutic sensitivity in heterogenous tumor subpopulations.

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Acknowledgements

The authors would like to thank A. Burçin Asutay for technical assistance during flow cytometry analysis.

Funding

This study was funded by the Scientific and Technological Research Council of Turkey (TUBITAK), Project No: 315S039.

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Authors and Affiliations

  1. Department of Genetics and Bioengineering, Faculty of Engineering and Architecture, Yeditepe University, Istanbul, Turkey

    Ezgi Avsar Abdik & Fikrettin Sahin

  2. Department of Molecular Biology and Genetics, Faculty of Engineering and Natural Sciences, Istanbul Sabahattin Zaim University, Istanbul, Turkey

    Hüseyin Abdik

  3. Department of Cellular and Molecular Medicine, Faculty of Health Sciences, Koç University, Istanbul, Turkey

    Duygu Turan

  4. Toxicology and Chemotherapy Unit, German Cancer Research Center, Heidelberg, Germany

    Martin R. Berger

  5. Department of Pharmacology and Clinical Pharmacology, Istinye University Faculty of Medicine, Topkapı Campus, Maltepe Neighbourhood, Teyyareci Sami St., No. 3, Zeytinburnu, Istanbul, Turkey

    Ferda Kaleagasioglu

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Contributions

EAA: Investigation, Methodology, Validation, Visualization, Writing—original draft. FK: Conceptualization, Methodology, Writing—original draft, Writing—review & editing, Project administration. HA: Investigation, Formal analysis. DT: Investigation. FS: Funding acquisition, Resources. Martin R. Berger: Resources, Writing—review & editing.

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Correspondence to Ferda Kaleagasioglu.

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Avsar Abdik, E., Abdik, H., Turan, D. et al. Dual Akt and Bcl-2 inhibition induces cell-type specific modulation of apoptotic and autophagic signaling in castration resistant prostate cancer cell lines. Mol Biol Rep 48, 7755–7765 (2021). https://doi.org/10.1007/s11033-021-06786-z

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