Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 Jun 25;25(13):6939.
doi: 10.3390/ijms25136939.

Mitochondrial Elongation and ROS-Mediated Apoptosis in Prostate Cancer Cells under Therapy with Apalutamide and Complex I Inhibitor

Valentin Baumgartner  1 , Dominik Schaer  2 , Holger Moch  3 , Souzan Salemi  1 , Daniel Eberli  1
Affiliations

Mitochondrial Elongation and ROS-Mediated Apoptosis in Prostate Cancer Cells under Therapy with Apalutamide and Complex I Inhibitor

Valentin Baumgartner et al. Int J Mol Sci. .

Abstract

Metabolic reprogramming and mitochondrial dynamics are pivotal in prostate cancer (PCa) progression and treatment resistance, making them essential targets for therapeutic intervention. In this study, we investigated the effects of the androgen receptor antagonist apalutamide (ARN) and the mitochondrial electron transport chain complex I inhibitor IACS-010759 (IACS) on the mitochondrial network architecture and dynamics in PCa cells. Treatment with ARN and/or IACS induced significant changes in mitochondrial morphology, particularly elongation, in androgen-sensitive PCa cells. Additionally, ARN and IACS modulated the mitochondrial fission and fusion processes, indicating a convergence of metabolic and androgen-signaling pathways in shaping mitochondrial function. Notably, the combination treatment with ARN and IACS resulted in increased apoptotic cell death and mitochondrial oxidative stress selectively in the androgen-sensitive PCa cells. Our findings highlight the therapeutic potential of targeting mitochondrial metabolism in prostate cancer and emphasize the need for further mechanistic understanding to optimize treatment strategies and improve patient outcomes.

Keywords: IACS-010759; apalutamide; mitochondria; oxidative stress; prostate cancer.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Mitochondrial network alterations in treated PCa cells. (A) Mitochondria were stained with TOM20 (red). Treatment with 25 µM apalutamide (ARN), 10 nM IACS-010759 (IACS) and Combo elongated mitochondria in LNCaP cells (top row). No morphological changes were observed in PC-3 cells upon drug treatment (bottom row). White boxed areas depict enlarged views of stained mitochondria. Scale bar: 50 µm. (B) TEM images visualize single mitochondria in LNCaP cells treated with DMSO, ARN, IACS and Combo. White arrows show representative mitochondria for each treatment condition. Scale bar: 1 µm. Quantification of n = 100 mitochondria for each treatment group with shape descriptors: area (C), perimeter (D), length (E) and roundness (F). Data are presented as means ± SEM. * p = 0.0458; ** p = 0.0030; *** p < 0.001; **** p < 0.0001. The uncropped TEM images are shown in Figure S1.
Figure 2
Figure 2
Treatment-induced changes in fission and fusion proteins. (A,B) Antibodies targeting OPA1 (A) and p-DRP1 (Ser616) (B) were quantified through automated Western blot analysis to determine changes in fusion and fission protein expression in PNT1A, LNCaP, C4-2 and PC-3 cells upon treatment with DMSO, ARN, IACS and Combo. The quantification of protein levels normalized to GAPDH is presented as means ± SEM (n = 3). * p < 0.05; ** p = 0.0034; *** p < 0.001; **** p < 0.0001. (C) Representative virtual immunoblots are shown for OPA1 and p-DRP1 (Ser616). Uncropped lanes are shown in Figures S2 and S3.
Figure 3
Figure 3
Androgen supplementation increases mitochondrial respiration and affects mitochondrial network. (A) Seahorse extracellular flux assay measured OCR in androgen-depleted (EtOH) and supplemented (DHT) LNCaP cells. Sequential addition of oligomycin, FCCP and rotenone + antimycin A were used to quantify basal respiration, ATP production and maximum respiration (B). (C) Cells were immunolabeled with TOM20 to visualize the mitochondrial network in EtOH-treated (top) and DHT-treated (bottom) LNCaP cells. (D) Protein levels with active fission marker p-DRP1 (Ser616) for LNCaP (left) and PC-3 cells (right), with or without DHT addition. GAPDH was used as a loading control. Quantification of p-DRP1 normalized to GAPDH for LNCaP (E) and PC-3 cells (F). (G) Immunostained LNCaP (top row) and PC-3 (bottom row) with p-DRP1 (red), with or without DHT addition. Nuclei were stained with DAPI (blue). Scale bar: 50 µm. Bar plots represent means ± SEM (n = 3). * p < 0.05; ** p = 0.0058. Uncropped Western blot lanes are shown in Figure S4.
Figure 4
Figure 4
Assessment of apoptosis in PCa cells. (A) Representative flow cytometry dot plots of cells stained with Annexin V/PI. PCa cell lines PNT1A, LNCaP, C4-2 and PC-3 were treated for 72 h with DMSO, ARN, IACS and Combo. The sum of Annexin V (+)/PI (−) and Annexin V (+)/PI (+) cells in the lower right and upper right corner was used to quantify apoptotic cells. (B) Bar plots represent three independent experiments of measurements of apoptotic cells for PNT1A, LNCaP, C4-2 and PC-3. Bar plots are shown as means ± SEM. * p = 0.0179; ** p < 0.01.
Figure 5
Figure 5
IACS and Combo increase mitochondrial ROS production in androgen-sensitive cells. (A) Oxidative stress in representative images of live cells labeled with MitoSOX (red) is shown. Scale bar: 200 µm. (B) Relative fluorescence intensity for MitoSOX was quantified for each cell line and treatment group (n = 3). Fluorescence intensity was normalized to DMSO control (100%). Bar plots represent means ± SEM. ** p < 0.01; *** p < 0.001; **** p < 0.0001.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Smith M.R., Saad F., Chowdhury S., Oudard S., Hadaschik B.A., Graff J.N., Olmos D., Mainwaring P.N., Lee J.Y., Uemura H., et al. Apalutamide Treatment and Metastasis-free Survival in Prostate Cancer. N. Engl. J. Med. 2018;378:1408–1418. doi: 10.1056/NEJMoa1715546. - DOI - PubMed
    1. Davis I.D., Martin A.J., Stockler M.R., Begbie S., Chi K.N., Chowdhury S., Coskinas X., Frydenberg M., Hague W.E., Horvath L.G., et al. Enzalutamide with Standard First-Line Therapy in Metastatic Prostate Cancer. N. Engl. J. Med. 2019;381:121–131. doi: 10.1056/NEJMoa1903835. - DOI - PubMed
    1. Fizazi K., Shore N., Tammela T.L., Ulys A., Vjaters E., Polyakov S., Jievaltas M., Luz M., Alekseev B., Kuss I., et al. Darolutamide in Nonmetastatic, Castration-Resistant Prostate Cancer. N. Engl. J. Med. 2019;380:1235–1246. doi: 10.1056/NEJMoa1815671. - DOI - PubMed
    1. Claessens F., Helsen C., Prekovic S., Van den Broeck T., Spans L., Van Poppel H., Joniau S. Emerging mechanisms of enzalutamide resistance in prostate cancer. Nat. Rev. Urol. 2014;11:712–716. doi: 10.1038/nrurol.2014.243. - DOI - PubMed
    1. Chandrasekar T., Yang J.C., Gao A.C., Evans C.P. Mechanisms of resistance in castration-resistant prostate cancer (CRPC) Transl. Androl. Urol. 2015;4:365–380. doi: 10.3978/j.issn.2223-4683.2015.05.02. - DOI - PMC - PubMed

MeSH terms