The Apoptotic and Anti-Warburg Effects of Brassinin in PC-3 Cells via Reactive Oxygen Species Production and the Inhibition of the c-Myc, SIRT1, and β-Catenin Signaling Axis
Abstract
:1. Introduction
2. Results
2.1. Cytotoxic Effect of Brassinin in Human Prostate Cancer Cells
2.2. Brassinin Induced Apoptosis in PC-3 Cells
2.3. Brassinin Effectively Suppressed the Expression of Glycolysis-Related Proteins in PC-3 Cells
2.4. Brassinin Reduces the Expression of SIRT1, β-Catenin, and c-Myc in PC-3 Cells
2.5. Effect of Brassinin on Interaction between SIRT1 and β-Catenin in PC-3 Cells
2.6. Brassinin Increased ROS Production in PC-3 Cells
2.7. ROS Inhibitor N-Acetyl-L-Cysteine (NAC), Disturbs Brassinin-Induced Apoptosis in PC-3 Cells
3. Discussion
4. Materials and Methods
4.1. Brassinin Preparation
4.2. Cell Culture
4.3. Cytotoxicity Assay
4.4. Colony Formation Assay
4.5. Cell Cycle Analysis
4.6. Terminal Deoxynucleotidyl Transferase dUTP Nick end Labeling (TUNEL) Assay
4.7. Western Blotting
4.8. Co-Immunoprecipitation
4.9. Measurement of ROS Generation
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
- Sung, H.; Ferlay, J.; Siegel, R.L.; Laversanne, M.; Soerjomataram, I.; Jemal, A.; Bray, F. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J. Clin. 2021, 71, 209–249. [Google Scholar] [CrossRef]
- Bergengren, O.; Pekala, K.R.; Matsoukas, K.; Fainberg, J.; Mungovan, S.F.; Bratt, O.; Bray, F.; Brawley, O.; Luckenbaugh, A.N.; Mucci, L.; et al. 2022 Update on Prostate Cancer Epidemiology and Risk Factors-A Systematic Review. Eur. Urol. 2023, 84, 191–206. [Google Scholar] [CrossRef] [PubMed]
- World Health Organization. Data Visualization Tools for Exploring the Global Cancer Burnden in 2020; World Health Organization: Geneva, Switzerland, 2020; Available online: https://gco.iarc.fr/ (accessed on 24 August 2023).
- Teo, M.Y.; Rathkopf, D.E.; Kantoff, P. Treatment of Advanced Prostate Cancer. Annu. Rev. Med. 2019, 70, 479–499. [Google Scholar] [CrossRef] [PubMed]
- Gourdin, T. Recent progress in treating advanced prostate cancer. Curr. Opin. Oncol. 2020, 32, 210–215. [Google Scholar] [CrossRef]
- Haberkorn, U.; Eder, M.; Kopka, K.; Babich, J.W.; Eisenhut, M. New Strategies in Prostate Cancer: Prostate-Specific Membrane Antigen (PSMA) Ligands for Diagnosis and Therapy. Clin. Cancer Res. 2016, 22, 9–15. [Google Scholar] [CrossRef]
- Eloy, J.O.; Ruiz, A.; de Lima, F.T.; Petrilli, R.; Raspantini, G.; Nogueira, K.A.B.; Santos, E.; de Oliveira, C.S.; Borges, J.C.; Marchetti, J.M.; et al. EGFR-targeted immunoliposomes efficiently deliver docetaxel to prostate cancer cells. Colloids Surf. B Biointerfaces 2020, 194, 111185. [Google Scholar] [CrossRef]
- Melegh, Z.; Oltean, S. Targeting Angiogenesis in Prostate Cancer. Int. J. Mol. Sci. 2019, 20, 2676. [Google Scholar] [CrossRef] [PubMed]
- Lee, I.I.; Kuznik, N.C.; Rottenberg, J.T.; Brown, M.; Cato, A.C.B. BAG1L: A promising therapeutic target for androgen receptor-dependent prostate cancer. J. Mol. Endocrinol. 2019, 62, R289–R299. [Google Scholar] [CrossRef]
- Schwartz, L.; Supuran, C.T.; Alfarouk, K.O. The Warburg Effect and the Hallmarks of Cancer. Anticancer Agents Med. Chem. 2017, 17, 164–170. [Google Scholar] [CrossRef]
- Dyshlovoy, S.A.; Pelageev, D.N.; Hauschild, J.; Borisova, K.L.; Kaune, M.; Krisp, C.; Venz, S.; Sabutskii, Y.E.; Khmelevskaya, E.A.; Busenbender, T.; et al. Successful Targeting of the Warburg Effect in Prostate Cancer by Glucose-Conjugated 1,4-Naphthoquinones. Cancers 2019, 11, 1690. [Google Scholar] [CrossRef]
- Song, C.; Zhang, J.; Liu, X.; Li, M.; Wang, D.; Kang, Z.; Yu, J.; Chen, J.; Pan, H.; Wang, H.; et al. PTEN loss promotes Warburg effect and prostate cancer cell growth by inducing FBP1 degradation. Front. Oncol. 2022, 12, 911466. [Google Scholar] [CrossRef] [PubMed]
- Graziano, F.; Ruzzo, A.; Giacomini, E.; Ricciardi, T.; Aprile, G.; Loupakis, F.; Lorenzini, P.; Ongaro, E.; Zoratto, F.; Catalano, V.; et al. Glycolysis gene expression analysis and selective metabolic advantage in the clinical progression of colorectal cancer. Pharmacogenom. J. 2017, 17, 258–264. [Google Scholar] [CrossRef]
- Yao, G.; Yin, J.; Wang, Q.; Dong, R.; Lu, J. Glypican-3 Enhances Reprogramming of Glucose Metabolism in Liver Cancer Cells. Biomed. Res. Int. 2019, 2019, 2560650. [Google Scholar] [CrossRef]
- Russo, G.I.; Asmundo, M.G.; Lo Giudice, A.; Trefiletti, G.; Cimino, S.; Ferro, M.; Lombardo, R.; De Nunzio, C.; Morgia, G.; Piombino, E.; et al. Is There a Role of Warburg Effect in Prostate Cancer Aggressiveness? Analysis of Expression of Enzymes of Lipidic Metabolism by Immunohistochemistry in Prostate Cancer Patients (DIAMOND Study). Cancers 2023, 15, 948. [Google Scholar] [CrossRef]
- Duffy, M.J.; O’Grady, S.; Tang, M.; Crown, J. MYC as a target for cancer treatment. Cancer Treat. Rev. 2021, 94, 102154. [Google Scholar] [CrossRef]
- Fleming, N.I.; Jorissen, R.N.; Mouradov, D.; Christie, M.; Sakthianandeswaren, A.; Palmieri, M.; Day, F.; Li, S.; Tsui, C.; Lipton, L.; et al. SMAD2, SMAD3 and SMAD4 mutations in colorectal cancer. Cancer Res. 2013, 73, 725–735. [Google Scholar] [CrossRef] [PubMed]
- Peng, Y.; Liu, J.; Wang, Z.; Cui, C.; Zhang, T.; Zhang, S.; Gao, P.; Hou, Z.; Liu, H.; Guo, J.; et al. Prostate-specific oncogene OTUD6A promotes prostatic tumorigenesis via deubiquitinating and stabilizing c-Myc. Cell Death Differ. 2022, 29, 1730–1743. [Google Scholar] [CrossRef] [PubMed]
- Dang, C.V.; Le, A.; Gao, P. MYC-induced cancer cell energy metabolism and therapeutic opportunities. Clin. Cancer Res. 2009, 15, 6479–6483. [Google Scholar] [CrossRef]
- Wu, Y.; Meng, X.; Huang, C.; Li, J. Emerging role of silent information regulator 1 (SIRT1) in hepatocellular carcinoma: A potential therapeutic target. Tumour Biol. 2015, 36, 4063–4074. [Google Scholar] [CrossRef]
- Wei, Z.; Xia, J.; Li, J.; Cai, J.; Shan, J.; Zhang, C.; Zhang, L.; Wang, T.; Qian, C.; Liu, L. SIRT1 promotes glucolipid metabolic conversion to facilitate tumor development in colorectal carcinoma. Int. J. Biol. Sci. 2023, 19, 1925–1940. [Google Scholar] [CrossRef]
- Murillo-Garzón, V.; Kypta, R. WNT signalling in prostate cancer. Nat. Rev. Urol. 2017, 14, 683–696. [Google Scholar] [CrossRef] [PubMed]
- Vallée, A.; Lecarpentier, Y.; Guillevin, R.; Vallée, J.N. Aerobic Glycolysis Hypothesis Through WNT/Beta-Catenin Pathway in Exudative Age-Related Macular Degeneration. J. Mol. Neurosci. 2017, 62, 368–379. [Google Scholar] [CrossRef] [PubMed]
- Vallée, A.; Lecarpentier, Y.; Vallée, J.N. The Key Role of the WNT/β-Catenin Pathway in Metabolic Reprogramming in Cancers under Normoxic Conditions. Cancers 2021, 13, 5557. [Google Scholar] [CrossRef] [PubMed]
- Lu, J.; Tan, M.; Cai, Q. The Warburg effect in tumor progression: Mitochondrial oxidative metabolism as an anti-metastasis mechanism. Cancer Lett. 2015, 356, 156–164. [Google Scholar] [CrossRef] [PubMed]
- Tuli, H.S.; Kaur, J.; Vashishth, K.; Sak, K.; Sharma, U.; Choudhary, R.; Behl, T.; Singh, T.; Sharma, S.; Saini, A.K.; et al. Molecular mechanisms behind ROS regulation in cancer: A balancing act between augmented tumorigenesis and cell apoptosis. Arch. Toxicol. 2023, 97, 103–120. [Google Scholar] [CrossRef]
- Termini, D.; Den Hartogh, D.J.; Jaglanian, A.; Tsiani, E. Curcumin against Prostate Cancer: Current Evidence. Biomolecules 2020, 10, 1536. [Google Scholar] [CrossRef]
- Traka, M.H.; Melchini, A.; Mithen, R.F. Sulforaphane and prostate cancer interception. Drug Discov. Today 2014, 19, 1488–1492. [Google Scholar] [CrossRef]
- Yang, F.; Song, L.; Wang, H.; Wang, J.; Xu, Z.; Xing, N. Quercetin in prostate cancer: Chemotherapeutic and chemopreventive effects, mechanisms and clinical application potential (Review). Oncol. Rep. 2015, 33, 2659–2668. [Google Scholar] [CrossRef]
- Samy, A.; Shah, D.; Shahagadkar, P.; Shah, H.; Munirathinam, G. Can diallyl trisulfide, a dietary garlic-derived compound, activate ferroptosis to overcome therapy resistance in prostate cancer? Nutr. Health 2022, 28, 207–212. [Google Scholar] [CrossRef]
- Tang, Y.; Chen, R.; Huang, Y.; Li, G.; Huang, Y.; Chen, J.; Duan, L.; Zhu, B.T.; Thrasher, J.B.; Zhang, X.; et al. Natural compound Alternol induces oxidative stress-dependent apoptotic cell death preferentially in prostate cancer cells. Mol. Cancer Ther. 2014, 13, 1526–1536. [Google Scholar] [CrossRef]
- Sherman, B.; Hernandez, A.M.; Alhado, M.; Menge, L.; Price, R.S. Silibinin Differentially Decreases the Aggressive Cancer Phenotype in an In Vitro Model of Obesity and Prostate Cancer. Nutr. Cancer 2020, 72, 333–342. [Google Scholar] [CrossRef]
- Jiang, C.; Lee, H.J.; Li, G.X.; Guo, J.; Malewicz, B.; Zhao, Y.; Lee, E.O.; Lee, H.J.; Lee, J.H.; Kim, M.S.; et al. Potent antiandrogen and androgen receptor activities of an Angelica gigas-containing herbal formulation: Identification of decursin as a novel and active compound with implications for prevention and treatment of prostate cancer. Cancer Res. 2006, 66, 453–463. [Google Scholar] [CrossRef] [PubMed]
- Kang, B.; Hwang, J.; Choi, H.S. Brassinin, a brassica-derived phytochemical, regulates monocyte-to-macrophage differentiation and inflammatory responses in human monocytes and murine macrophages. J. Pharm. Pharmacol. 2020, 72, 1245–1255. [Google Scholar] [CrossRef] [PubMed]
- Hong, T.; Ham, J.; Song, J.; Song, G.; Lim, W. Brassinin Inhibits Proliferation in Human Liver Cancer Cells via Mitochondrial Dysfunction. Cells 2021, 10, 332. [Google Scholar] [CrossRef]
- Yang, M.H.; Baek, S.H.; Ha, I.J.; Um, J.Y.; Ahn, K.S. Brassinin enhances the anticancer actions of paclitaxel by targeting multiple signaling pathways in colorectal cancer cells. Phytother. Res. 2021, 35, 3875–3885. [Google Scholar] [CrossRef]
- Kim, S.M.; Oh, E.Y.; Lee, J.H.; Nam, D.; Lee, S.G.; Lee, J.; Kim, S.H.; Shim, B.S.; Ahn, K.S. Brassinin Combined with Capsaicin Enhances Apoptotic and Anti-metastatic Effects in PC-3 Human Prostate Cancer Cells. Phytother. Res. 2015, 29, 1828–1836. [Google Scholar] [CrossRef]
- Gu, Y.; Becker, V.; Qiu, M.; Tang, T.; Ampofo, E.; Menger, M.D.; Laschke, M.W. Brassinin Promotes the Degradation of Tie2 and FGFR1 in Endothelial Cells and Inhibits Triple-Negative Breast Cancer Angiogenesis. Cancers 2022, 14, 3540. [Google Scholar] [CrossRef]
- Yang, M.H.; Lee, J.H.; Ko, J.H.; Jung, S.H.; Sethi, G.; Ahn, K.S. Brassinin Represses Invasive Potential of Lung Carcinoma Cells through Deactivation of PI3K/Akt/mTOR Signaling Cascade. Molecules 2019, 24, 1584. [Google Scholar] [CrossRef]
- Gerhäuser, C.; You, M.; Liu, J.; Moriarty, R.M.; Hawthorne, M.; Mehta, R.G.; Moon, R.C.; Pezzuto, J.M. Cancer chemopreventive potential of sulforamate, a novel analogue of sulforaphane that induces phase 2 drug-metabolizing enzymes. Cancer Res. 1997, 57, 272–278. [Google Scholar] [PubMed]
- Kim, S.M.; Park, J.H.; Kim, K.D.; Nam, D.; Shim, B.S.; Kim, S.H.; Ahn, K.S.; Choi, S.H.; Ahn, K.S. Brassinin induces apoptosis in PC-3 human prostate cancer cells through the suppression of PI3K/Akt/mTOR/S6K1 signaling cascades. Phytother. Res. 2014, 28, 423–431. [Google Scholar] [CrossRef]
- Ware, J.L.; Paulson, D.F.; Mickey, G.H.; Webb, K.S. Spontaneous metastasis of cells of the human prostate carcinoma cell line PC-3 in athymic nude mice. J. Urol. 1982, 128, 1064–1067. [Google Scholar] [CrossRef] [PubMed]
- Camby, I.; Etievant, C.; Petein, M.; Dedecker, R.; van Velthoven, R.; Danguy, A.; Pasteels, J.L.; Kiss, R. Influence of culture media on the morphological differentiation of the PC-3 and DU145 prostatic neoplastic cell lines. Prostate 1994, 24, 187–196. [Google Scholar] [CrossRef] [PubMed]
- Wen, X.; Lin, Z.Q.; Liu, B.; Wei, Y.Q. Caspase-mediated programmed cell death pathways as potential therapeutic targets in cancer. Cell Prolif. 2012, 45, 217–224. [Google Scholar] [CrossRef] [PubMed]
- Lizard, G.; Miguet, C.; Gueldry, S.; Monier, S.; Gambert, P. Flow cytometry measurement of DNA fragmentation in the course of cell death via apoptosis. New techniques for evaluation of DNA status for the pathologist. Ann. Pathol. 1997, 17, 61–66. [Google Scholar]
- Vaupel, P.; Schmidberger, H.; Mayer, A. The Warburg effect: Essential part of metabolic reprogramming and central contributor to cancer progression. Int. J. Radiat. Biol. 2019, 95, 912–919. [Google Scholar] [CrossRef]
- Goetzman, E.S.; Prochownik, E.V. The Role for Myc in Coordinating Glycolysis, Oxidative Phosphorylation, Glutaminolysis, and Fatty Acid Metabolism in Normal and Neoplastic Tissues. Front. Endocrinol. 2018, 9, 129. [Google Scholar] [CrossRef]
- Chen, Y.; Yang, H.; Chen, S.; Lu, Z.; Li, B.; Jiang, T.; Xuan, M.; Ye, R.; Liang, H.; Liu, X.; et al. SIRT1 regulated hexokinase-2 promoting glycolysis is involved in hydroquinone-enhanced malignant progression in human lymphoblastoid TK6 cells. Ecotoxicol. Environ. Saf. 2022, 241, 113757. [Google Scholar] [CrossRef]
- Fan, Q.; Yang, L.; Zhang, X.; Ma, Y.; Li, Y.; Dong, L.; Zong, Z.; Hua, X.; Su, D.; Li, H.; et al. Autophagy promotes metastasis and glycolysis by upregulating MCT1 expression and Wnt/β-catenin signaling pathway activation in hepatocellular carcinoma cells. J. Exp. Clin. Cancer Res. 2018, 37, 9. [Google Scholar] [CrossRef]
- Fang, Y.; Shen, Z.Y.; Zhan, Y.Z.; Feng, X.C.; Chen, K.L.; Li, Y.S.; Deng, H.J.; Pan, S.M.; Wu, D.H.; Ding, Y. CD36 inhibits β-catenin/c-myc-mediated glycolysis through ubiquitination of GPC4 to repress colorectal tumorigenesis. Nat. Commun. 2019, 10, 3981. [Google Scholar] [CrossRef]
- Zhou, Y.; Song, T.; Peng, J.; Zhou, Z.; Wei, H.; Zhou, R.; Jiang, S.; Peng, J. SIRT1 suppresses adipogenesis by activating Wnt/β-catenin signaling in vivo and in vitro. Oncotarget 2016, 7, 77707–77720. [Google Scholar] [CrossRef]
- Moloney, J.N.; Cotter, T.G. ROS signalling in the biology of cancer. Semin. Cell Dev. Biol. 2018, 80, 50–64. [Google Scholar] [CrossRef] [PubMed]
- Castelli, S.; Ciccarone, F.; Tavian, D.; Ciriolo, M.R. ROS-dependent HIF1α activation under forced lipid catabolism entails glycolysis and mitophagy as mediators of higher proliferation rate in cervical cancer cells. J. Exp. Clin. Cancer Res. 2021, 40, 94. [Google Scholar] [CrossRef] [PubMed]
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Kwon, H.H.; Ahn, C.-H.; Lee, H.-J.; Sim, D.Y.; Park, J.E.; Park, S.-Y.; Kim, B.; Shim, B.-S.; Kim, S.-H. The Apoptotic and Anti-Warburg Effects of Brassinin in PC-3 Cells via Reactive Oxygen Species Production and the Inhibition of the c-Myc, SIRT1, and β-Catenin Signaling Axis. Int. J. Mol. Sci. 2023, 24, 13912. https://doi.org/10.3390/ijms241813912
Kwon HH, Ahn C-H, Lee H-J, Sim DY, Park JE, Park S-Y, Kim B, Shim B-S, Kim S-H. The Apoptotic and Anti-Warburg Effects of Brassinin in PC-3 Cells via Reactive Oxygen Species Production and the Inhibition of the c-Myc, SIRT1, and β-Catenin Signaling Axis. International Journal of Molecular Sciences. 2023; 24(18):13912. https://doi.org/10.3390/ijms241813912
Chicago/Turabian StyleKwon, Hyeon Hee, Chi-Hoon Ahn, Hyo-Jung Lee, Deok Yong Sim, Ji Eon Park, Su-Yeon Park, Bonglee Kim, Bum-Sang Shim, and Sung-Hoon Kim. 2023. "The Apoptotic and Anti-Warburg Effects of Brassinin in PC-3 Cells via Reactive Oxygen Species Production and the Inhibition of the c-Myc, SIRT1, and β-Catenin Signaling Axis" International Journal of Molecular Sciences 24, no. 18: 13912. https://doi.org/10.3390/ijms241813912