In Silico and In Vitro Investigation of Apoptosis-Mediated Antiproliferative Activity of Capsaicin and Alpha-Lipoic Acid Against Prostate Cancer Cells
Abstract
1. Introduction
2. Materials and Methods
2.1. Reagents
2.2. Cell Culture
2.3. Cell Viability Assay
2.4. Western Blot Analysis
2.5. Molecular Docking
2.6. Drug-likeness and Pharmacokinetic Analysis
2.7. Bioavailability Radar
2.8. Statistical Analysis
3. Results
3.1. Effects of Capsaicin and ALA on Cell Viability
3.2. Effects of Capsaicin and ALA on Protein Expression
3.3. Molecular Docking Analysis
3.4. Drug-likeness and Pharmacokinetic Properties
3.5. Bioavailability Radar
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
- Vietri, M.T.; D’elia, G.; Caliendo, G.; Resse, M.; Casamassimi, A.; Passariello, L.; Albanese, L.; Cioffi, M.; Molinari, A.M. Hereditary prostate cancer: Genes related, target therapy and prevention. Int. J. Mol. Sci. 2021, 22, 3753. [Google Scholar] [CrossRef]
- Bai, B.; Chen, Q.; Jing, R.; He, X.; Wang, H.; Ban, Y.; Ye, Q.; Xu, W.; Zheng, C. Molecular basis of prostate cancer and natural products as potential chemotherapeutic and chemopreventive agents. Front. Pharmacol. 2021, 12, 738235. [Google Scholar] [CrossRef] [PubMed]
- Licitra, F.; Giovannelli, P.; Di Donato, M.; Monaco, A.; Galasso, G.; Migliaccio, A.; Castoria, G. New insights and emerging therapeutic approaches in prostate cancer. Front. Endocrinol. 2022, 13, 840787. [Google Scholar] [CrossRef] [PubMed]
- Parnes, H.L.; House, M.G.; Tangrea, J.A. Prostate cancer prevention: Agent development strategies. In Recent Results in Cancer Research; Springer: Berlin/Heidelberg, Germany, 2014; Volume 202, pp. 121–131. [Google Scholar] [CrossRef]
- Du, J.; Tang, X.L. Natural products against cancer: A comprehensive bibliometric study of the research projects, publications, patents and drugs. J. Cancer Res. Ther. 2014, 10, 27–37. [Google Scholar] [CrossRef]
- Hashem, S.; Ali, T.A.; Akhtar, S.; Nisar, S.; Sageena, G.; Ali, S.; Al-Mannai, S.; Therachiyil, L.; Mir, R.; Elfaki, I.; et al. Targeting cancer signaling pathways by natural products: Exploring promising anti-cancer agents. Biomed. Pharmacother. 2022, 150, 113054. [Google Scholar] [CrossRef]
- Wang, Y.; Zhong, J.; Bai, J.; Tong, R.; An, F.; Jiao, P.; He, L.; Zeng, D.; Long, E.; Yan, J.; et al. The application of natural products in cancer therapy by targeting apoptosis pathways. Curr. Drug Metab. 2018, 19, 739–749. [Google Scholar] [CrossRef]
- Cavalcante, G.C.; Schaan, A.P.; Cabral, G.F.; Santana-Da-Silva, M.N.; Pinto, P.; Vidal, A.F.; Ribeiro-Dos-Santos, Â. A cell’s fate: An overview of the molecular biology and genetics of apoptosis. Int. J. Mol. Sci. 2019, 20, 4133. [Google Scholar] [CrossRef] [PubMed]
- Clark, R.; Lee, S.H. Anticancer properties of capsaicin against human cancer. Anticancer Res. 2016, 36, 837–843. [Google Scholar]
- Jun, H.-S.; Park, T.; Lee, C.K.; Kang, M.K.; Park, M.S.; Kang, H.I.; Surh, Y.-J.; Kim, O.H. Capsaicin induced apoptosis of B16-F10 melanoma cells through down-regulation of Bcl-2. Food Chem. Toxicol. 2007, 45, 708–715. [Google Scholar] [CrossRef]
- Salehi, B.; Berkay Yılmaz, Y.; Antika, G.; Boyunegmez Tumer, T.; Fawzi Mahomoodally, M.; Lobine, D.; Akram, M.; Riaz, M.; Capanoglu, E.; Sharopov, F.; et al. Insights on the use of α-lipoic acid for therapeutic purposes. Biomolecules 2019, 9, 356. [Google Scholar] [CrossRef]
- Zonooz, S.R.; Hasani, M.; Morvaridzadeh, M.; Pizarro, A.B.; Heydari, H.; Yosaee, S.; Rezamand, G.; Heshmati, J. Effect of alpha-lipoic acid on oxidative stress parameters: A systematic review and meta-analysis. J. Funct. Foods 2021, 87, 10477. [Google Scholar] [CrossRef]
- Shi, D.Y.; Liu, H.L.; Stern, J.S.; Yu, P.Z.; Liu, S.L. Alpha-lipoic acid induces apoptosis in hepatoma cells via the PTEN/Akt pathway. FEBS Lett. 2008, 582, 1667–1671. [Google Scholar] [CrossRef]
- Banerjee, P.; Ulker, O.C. Combinative ex vivo studies and in silico models ProTox-II for investigating the toxicity of chemicals used mainly in cosmetic products. Toxicol. Mech. Methods 2022, 32, 542–548. [Google Scholar] [CrossRef] [PubMed]
- Adki, K.M.; Murugesan, S.; Kulkarni, Y.A. In silico and in vivo toxicological evaluation of paeonol. Chem. Biodivers. 2020, 17, e2000422. [Google Scholar] [CrossRef] [PubMed]
- Liu, Y.; Yang, X.; Gan, J.; Chen, S.; Xiao, Z.-X.; Cao, Y. CB-Dock2: Improved protein-ligand blind docking by integrating cavity detection, docking and homologous template fitting. Nucleic Acids Res. 2022, 50, 159–164. [Google Scholar] [CrossRef]
- Ashraf, H.; Dilshad, E.; Afsar, T.; Almajwal, A.; Shafique, H.; Razak, S. Molecular screening of bioactive compounds of garlic for therapeutic effects against COVID-19. Biomedicines 2023, 11, 643. [Google Scholar] [CrossRef]
- Sympli, H.D. Estimation of drug-likeness properties of GC-MS separated bioactive compounds in rare medicinal pleione maculata using molecular docking technique and SwissADME in silico tools. Netw. Model. Anal. Health Inform. Bioinform. 2021, 10, 14. [Google Scholar] [CrossRef]
- Daina, A.; Michielin, O.; Zoete, V. SwissADME: A free web tool to evaluate pharmacokinetics, drug-likeness and medicinal chemistry friendliness of small molecules. Sci. Rep. 2017, 7, 42717. [Google Scholar] [CrossRef]
- Ranjan, A.; Ramachandran, S.; Gupta, N.; Kaushik, I.; Wright, S.; Srivastava, S.; Das, H.; Srivastava, S.; Prasad, S.; Srivastava, S.K. Role of phytochemicals in cancer prevention. Int. J. Mol. Sci. 2019, 20, 4981. [Google Scholar] [CrossRef]
- Sánchez, B.G.; Bort, A.; Mateos-Gómez, P.A.; Rodríguez-Henche, N.; Díaz-Laviada, I. Combination of the natural product capsaicin and docetaxel synergistically kills human prostate cancer cells through the metabolic regulator AMP-activated kinase. Cancer Cell Int. 2019, 19, 54. [Google Scholar] [CrossRef] [PubMed]
- Zhu, M.; Yu, X.; Zheng, Z.; Huang, J.; Yang, X.; Shi, H. Capsaicin suppressed activity of prostate cancer stem cells by inhibition of Wnt/β-catenin pathway. Phytother. Res. 2020, 34, 817–824. [Google Scholar] [CrossRef] [PubMed]
- Lewinska, A.; Jarosz, P.; Czech, J.; Rzeszutek, I.; Bielak-Zmijewska, A.; Grabowska, W.; Wnuk, M. Capsaicin-induced genotoxic stress does not promote apoptosis in A549 human lung and DU145 prostate cancer cells. Mutat. Res. Genet. Toxicol. 2015, 779, 23–34. [Google Scholar] [CrossRef]
- Bort, A.; Spínola, E.; Rodríguez-Henche, N.; Díaz-Laviada, I. Capsaicin exerts synergistic antitumor effect with sorafenib in hepatocellular carcinoma cells through AMPK activation. Oncotarget 2017, 8, 87684–87698. [Google Scholar] [CrossRef]
- Wang, Y.; Deng, X.; Yu, C.; Zhao, G.; Zhou, J.; Zhang, G.; Li, M.; Jiang, D.; Quan, Z.; Zhang, Y. Synergistic inhibitory effects of capsaicin combined with cisplatin on human osteosarcoma in culture and in xenografts. J. Exp. Clin. Cancer Res. 2018, 37, 251. [Google Scholar] [CrossRef]
- Clark, R.; Lee, J.; Lee, S.H. Synergistic anticancer activity of capsaicin and 3,3′-diindolylmethane in human colorectal cancer. J. Agric. Food Chem. 2015, 63, 4297–4304. [Google Scholar] [CrossRef]
- Zhang, R.; Humphreys, I.; Sahu, R.P.; Shi, Y.; Srivastava, S.K. In vitro and in vivo induction of apoptosis by capsaicin in pancreatic cancer cells is mediated through ROS generation and mitochondrial death pathway. Apoptosis 2008, 13, 1465–1478. [Google Scholar] [CrossRef]
- Bossio, S.; Perri, A.; Gallo, R.; De Bartolo, A.; Rago, V.; La Russa, D.; Di Dio, M.; La Vignera, S.; Calogero, A.E.; Vitale, G.; et al. Alpha-lipoic acid reduces cell growth, inhibits autophagy, and counteracts prostate cancer cell migration and invasion: Evidence from in vitro studies. Int. J. Mol. Sci. 2023, 24, 17111. [Google Scholar] [CrossRef]
- Simbula, G.; Columbano, A.; Ledda-Columbano, G.M.; Sanna, L.; Deidda, M.; Diana, A.; Pibiri, M. Increased ROS generation and p53 activation in alpha-lipoic acid-induced apoptosis of hepatoma cells. Apoptosis 2007, 12, 113–123. [Google Scholar] [CrossRef]
- Na, M.H.; Seo, E.Y.; Kim, W.K. Effects of alpha-lipoic acid on cell proliferation and apoptosis in MDA-MB-231 human breast cells. Nutr. Res. Pract. 2009, 3, 265–271. [Google Scholar] [CrossRef] [PubMed]
- Carneiro, B.A.; El-Deiry, W.S. Targeting apoptosis in cancer therapy. Nat. Rev. Clin. Oncol. 2020, 17, 395–417. [Google Scholar] [CrossRef] [PubMed]
- Pramanik, K.C.; Fofaria, N.M.; Gupta, P.; Ranjan, A.; Kim, S.-H.; Srivastava, S.K. Inhibition of β-catenin signaling suppresses pancreatic tumor growth by disrupting nuclear β-catenin/TCF-1 complex: Critical role of STAT-3. Oncotarget 2015, 6, 11561–11574. [Google Scholar] [CrossRef]
- Bley, K.; Boorman, G.; Mohammad, B.; McKenzie, D.; Babbar, S. A comprehensive review of the carcinogenic and anticarcinogenic potential of capsaicin. Toxicol. Pathol. 2012, 40, 847–873. [Google Scholar] [CrossRef] [PubMed]
- Pramanik, K.C.; Srivastava, S.K. Apoptosis signal-regulating kinase 1-thioredoxin complex dissociation by capsaicin causes pancreatic tumor growth suppression by inducing apoptosis. Antioxid. Redox Signal. 2012, 17, 1417–1432. [Google Scholar] [CrossRef]
- Sánchez, A.M.; Sánchez, M.G.; Malagarie-Cazenave, S.; Olea, N.; Díaz-Laviada, I. Induction of apoptosis in prostate tumor PC-3 cells and inhibition of xenograft prostate tumor growth by the vanilloid capsaicin. Apoptosis 2006, 11, 89–99. [Google Scholar] [CrossRef] [PubMed]
- Sánchez, B.G.; Bort, A.; Mora-Rodríguez, J.M.; Díaz-Laviada, I. The natural chemotherapeutic capsaicin activates AMPK through LKB1 kinase and TRPV1 receptors in prostate cancer cells. Pharmaceutics 2022, 14, 329. [Google Scholar] [CrossRef]
- Kishi, H.; Igawa, M.; Kikuno, N.; Yoshino, T.; Urakami, S.; Shiina, H. Expression of the survivin gene in prostate cancer: Correlation with clinicopathological characteristics, proliferative activity and apoptosis. J. Urol. 2004, 171, 1855–1860. [Google Scholar] [CrossRef]
- Liu, Z.; Ding, Y.; Ye, N.; Wild, C.; Chen, H.; Zhou, J. Direct activation of bax protein for cancer therapy. Med. Res. Rev. 2016, 36, 313–341. [Google Scholar] [CrossRef] [PubMed]
- Chen, M.; Xiao, C.; Jiang, W.; Yang, W.; Qin, Q.; Tan, Q.; Lian, B.; Liang, Z.; Wei, C. Capsaicin inhibits proliferation and induces apoptosis in breast cancer by down-regulating FBI-1-mediated NF-κB pathway. Drug Des. Dev. Ther. 2021, 15, 125–140. [Google Scholar] [CrossRef]
- Liu, X.; Gao, R.; Dong, Y.; Gao, L.; Zhao, Y.; Zhao, L.; Zhao, X.; Zhang, H. Survivin gene silencing sensitizes prostate cancer cells to selenium growth inhibition. BMC Cancer 2010, 10, 418. [Google Scholar] [CrossRef]
- Ceylanlı, D.; Şehirli, A.Ö.; Gençosman, S.; Teralı, K.; Şah, H.; Gülmez, N.; Sayıner, S. Protective effects of alpha-lipoic acid against 5-fluorouracil-induced gastrointestinal mucositis in rats. Antioxidants 2022, 11, 1930. [Google Scholar] [CrossRef]
- Attia, M.; Essa, E.A.; Zaki, R.M.; Elkordy, A.A. An overview of the antioxidant effects of ascorbic acid and alpha lipoic acid (in liposomal forms) as adjuvant in cancer treatment. Antioxidants 2020, 9, 359. [Google Scholar] [CrossRef] [PubMed]






| Protein | Ligand | Vina Score (kcal/mol) | Cavity Volume (Å3) | Pocket RMSD | Center (x, y, z) | Docking Size (x, y, z) | Contact Residues |
|---|---|---|---|---|---|---|---|
| Survivin | Capsaicin | −6.6 | 226 | 0.88 | 57, −17, 35 | 24, 24, 24 | Chain A: PHE13 LEU14 LYS15 ARG18 ALA39 GLU40 ALA41 PHE58 PHE59 ILE74 HIS77 LYS78 ALA85 PHE86 LEU87 SER88 VAL89 LYS90 LYS91 GLN92 PHE93 LEU96 PHE101 LEU104 |
| Survivin | ALA | −4.9 | 565 | 1.20 | 29,1, 17 | 18, 18, 18 | Chain A: PHE13 LEU14 LYS15 ASP16 ARG18 ALA39 GLU40 ALA41 PHE58 ILE74 HIS77 LYS78 PHE86 LEU87 VAL89 LYS90 LYS91 GLN92 PHE93 GLU94 LEU96 PHE101 LEU104 |
| Bax | Capsaicin | −6.7 | 344 | 0.92 | 17, −9, 14 | 24, 24, 24 | Chain A: GLN28 ILE31 GLN32 ALA35 GLU41 ALA42 PRO43 LEU45 ALA46 LEU47 ASP48 PRO49 VAL121 LEU125 PRO130 GLU131 ILE133 ARG134 MET137 |
| Bax | ALA | −4.7 | 306 | 1.55 | 49, 9, 19 | 18, 18, 18 | Chain B: GLN28 ILE31 GLN32 ALA35 GLU41 ALA42 PRO43 LEU45 ALA46 LEU47 LEU125 PRO130 GLU131 ILE133 ARG134 MET137 |
| Ligand | Molecular Formula | Molecular Weight (g/mol) | Rotatable Bonds | H Bond Acceptors | H Bond Donors | LogPO/W | GI Absorption | Water Solubility |
|---|---|---|---|---|---|---|---|---|
| ALA | C8H14O2S2 | 206.3 | 5.0 | 2.0 | 1.0 | 1.851 | LOW | SOLUBLE |
| Capsaicin | C18H27NO3 | 305.4 | 10.0 | 4.0 | 2.0 | 3.041 | LOW | LOW |
| Ligand | Lipinski Rule | Pfizer Rule | GSK Rule | Golden Triangle |
|---|---|---|---|---|
| ALA | Accepted | Accepted | Accepted | Accepted |
| Capsaicin | Accepted | Rejected | Accepted | Accepted |
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Meral, O.; Balkan, B.M.; Bestil, D.N.; Sayiner, S.; Ceylanli, D.; Uyurca, S.; Pehlivan, S.; Ozkurt, G.; Kismali, G.; Sel, T. In Silico and In Vitro Investigation of Apoptosis-Mediated Antiproliferative Activity of Capsaicin and Alpha-Lipoic Acid Against Prostate Cancer Cells. Curr. Issues Mol. Biol. 2026, 48, 376. https://doi.org/10.3390/cimb48040376
Meral O, Balkan BM, Bestil DN, Sayiner S, Ceylanli D, Uyurca S, Pehlivan S, Ozkurt G, Kismali G, Sel T. In Silico and In Vitro Investigation of Apoptosis-Mediated Antiproliferative Activity of Capsaicin and Alpha-Lipoic Acid Against Prostate Cancer Cells. Current Issues in Molecular Biology. 2026; 48(4):376. https://doi.org/10.3390/cimb48040376
Chicago/Turabian StyleMeral, Ogunc, Burcu Menekse Balkan, Dilek Nur Bestil, Serkan Sayiner, Deniz Ceylanli, Sare Uyurca, Sinem Pehlivan, Guzin Ozkurt, Gorkem Kismali, and Tevhide Sel. 2026. "In Silico and In Vitro Investigation of Apoptosis-Mediated Antiproliferative Activity of Capsaicin and Alpha-Lipoic Acid Against Prostate Cancer Cells" Current Issues in Molecular Biology 48, no. 4: 376. https://doi.org/10.3390/cimb48040376
APA StyleMeral, O., Balkan, B. M., Bestil, D. N., Sayiner, S., Ceylanli, D., Uyurca, S., Pehlivan, S., Ozkurt, G., Kismali, G., & Sel, T. (2026). In Silico and In Vitro Investigation of Apoptosis-Mediated Antiproliferative Activity of Capsaicin and Alpha-Lipoic Acid Against Prostate Cancer Cells. Current Issues in Molecular Biology, 48(4), 376. https://doi.org/10.3390/cimb48040376

