Home Life Sciences Diselenide-derivative of 3-pyridinol targets redox enzymes leading to cell cycle deregulation and apoptosis in A549 cells
Article
Licensed
Unlicensed Requires Authentication

Diselenide-derivative of 3-pyridinol targets redox enzymes leading to cell cycle deregulation and apoptosis in A549 cells

  • Vishwa V. Gandhi , Subhash C. Bihani , Prasad P. Phadnis and Amit Kunwar ORCID logo EMAIL logo
Published/Copyright: August 25, 2022
Biological Chemistry
From the journal Biological Chemistry Volume 403 Issue 10

Abstract

The aim of present study was to understand the mechanism of action of 2,2′-diselenobis(3-pyridinol) or DISPOL in human lung cancer (A549) cells. A549 cells were treated with 10 µM (∼IC50) of DISPOL for varying time points to corelate the intracellular redox changes with its cytotoxic effect. The results indicated that DISPOL treatment led to a time dependant decrease in the basal level of reactive oxygen species (ROS). Additionally, DISPOL treatment elevated the ratio of reduced (GSH) and oxidised (GSSG) glutathione by upregulating gamma-glutamylcysteine ligase (γ-GCL) involved in GSH biosynthesis and inhibiting the activities of redox enzymes responsible for GSH utilization and recycling, such as glutathione-S-transferase (GST) and glutathione reductase (GR). Molecular docking analysis suggests putative interactions of DISPOL with GST and GR which could account for its inhibitory effect on these enzymes. Further, DISPOL induced reductive environment preceded G1 arrest and apoptosis as evidenced by decreased expression of cell cycle genes (Cyclin D1 and Cyclin E1) and elevation of p21 and apoptotic markers (cleaved caspase 3 and cleaved PARP). The combinatorial experiments involving DISPOL and redox modulatory agents such as N-acetylcysteine (NAC) and buthionine sulfoximine (BSO) indeed confirmed the role of reductive stress in DISPOL-induced cell death. Finally, Lipinski’s rule suggests attributes of drug likeness in DISPOL. Taken together, DISPOL exhibits a novel mechanism of reductive stress-mediated cell death in A549 cells that warrants future exploration as anticancer agent.

Keywords: apoptosis; cell cycle arrest; organodiselenides; reactive oxygen species; redox modulation
Corresponding author: Amit Kunwar, Radiation and Photochemistry Division, Bhabha Atomic Research Centre, Mumbai 400085, India; and Homi Bhabha National Institute, Anushaktinagar, Mumbai 400094, India, E-mail:

Acknowledgments

The authors acknowledge Dr. Awadhesh Kumar, Head, RPC Division BARC and Dr. A. K. Tyagi, Associate Director, Chemistry Group, BARC for their support and encouragement.

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: None declared.

  3. Conflict of interest statement: The authors declare no conflicts of interest regarding this article.

References

Álvarez-Pérez, M., Ali, W., Marć, M.A., Handzlik, J., and Domínguez-Álvarez, E. (2018). Selenides and diselenides: a review of their anticancer and chemopreventive activity. Molecules 23: 628, https://doi.org/10.3390/molecules23030628.Search in Google Scholar

Brigelius-Flohé, R. (1999). Tissue-specific functions of individual glutathione peroxidases. Free Radic. Biol. Med. 27: 951–965, https://doi.org/10.1016/s0891-5849(99)00173-2.Search in Google Scholar

Chaudhari, K.R., Kunwar, A., Bhuvanesh, N., and Dey, S. (2020). Synthesis and anti-proliferative activities of amine capped Pd and Pt macrocycles of 4, 4’-dipyridylselenides. New J. Chem. 44: 7329–7337, https://doi.org/10.1039/c9nj06052a.Search in Google Scholar

Circu, M.L. and Aw, T.Y. (2010). Reactive oxygen species, cellular redox systems, and apoptosis. Free Radic. Biol. Med. 48: 749–762, https://doi.org/10.1016/j.freeradbiomed.2009.12.022.Search in Google Scholar

Ding, L., Cao, J., Lin, W., Chen, H., Xiong, X., Ao, H., Yu, M., Lin, J., and Cui, Q. (2020). The roles of cyclin-dependent kinases in cell-cycle progression and therapeutic strategies in human breast cancer. Int. J. Mol. Sci. 21: 1960, https://doi.org/10.3390/ijms21061960.Search in Google Scholar

Eruslanov, E. and Kusmartsev, S. (2010). Identification of ROS using oxidized DCFDA and flow-cytometry. Methods Mol. Biol. 594: 57–72.10.1007/978-1-60761-411-1_4Search in Google Scholar

Gandhi, V.V., Phadnis, P.P., and Kunwar, A. (2020). 2, 2’-Dipyridyl diselenide (Py2Se2) induces G1 arrest and apoptosis in human lung carcinoma (A549) cells through ROS scavenging and reductive stress. Metallomics 12: 1253–1266, https://doi.org/10.1039/d0mt00106f.Search in Google Scholar

Gandhi, V.V., Gandhi, K.A., Kumbhare, L.B., Goda, J.S., Gota, V., Priyadarsini, K.I., and Kunwar, A. (2021). 3, 3’-Diselenodipropionic acid (DSePA) induces reductive stress in A549 cells triggering p53-independent apoptosis: a novel mechanism for diselenides. Free Radic. Biol. Med. 175: 1–17, https://doi.org/10.1016/j.freeradbiomed.2021.08.017.Search in Google Scholar

Ganther, H.E. (1999). Selenium metabolism, selenoproteins and mechanisms of cancer prevention: complexities with thioredoxin reductase. Carcinogenesis 20: 1657–1666, https://doi.org/10.1093/carcin/20.9.1657.Search in Google Scholar

Habig, W.H., Pabst, M.J., and Jakoby, W.B. (1974). Glutathione-S-transferases. The first enzymatic step in mercapturic acid formation. J. Biol. Chem. 249: 7130–7139, https://doi.org/10.1016/s0021-9258(19)42083-8.Search in Google Scholar

Harper, J.W., Elledge, S.J., Keyomarsi, K., Dynlacht, B., Tsai, L.H., Zhang, P., Dobrowolski, S., Bai, C., Connell-Crowley, L., and Swindell, E. (1995). Inhibition of cyclin-dependent kinases by p21. Mol. Biol. Cell 6: 387–400, https://doi.org/10.1091/mbc.6.4.387.Search in Google Scholar

Hissin, P.J. and Hilf, R. (1976). A fluorometric method for determination of oxidized and reduced glutathione in tissues. Anal. Biochem. 74: 214–226, https://doi.org/10.1016/0003-2697(76)90326-2.Search in Google Scholar

Hodage, A., Prabhu, C., Phadnis, P., Wadawale, A., Priyadarsini, I., and Jain, V.K. (2012). Synthesis, characterization, structures and GPx mimicking activity of pyridyl and pyrimidyl based organoselenium compounds. J. Organomet. Chem. 720: 19–25, https://doi.org/10.1016/j.jorganchem.2012.08.035.Search in Google Scholar

Ishikawa, K., Ishii, H., and Saito, T. (2006). DNA damage-dependent cell cycle checkpoints and genomic stability. DNA Cell Biol. 25: 406–411, https://doi.org/10.1089/dna.2006.25.406.Search in Google Scholar

Kong, K.H., Takasu, K., Inoue, H., and Takahashi, K. (1992). Tyrosine-7 in human class Pi glutathione S-transferase is important for lowering the pKa of the thiol group of glutathione in the enzyme-glutathione complex. Biochem. Biophys. Res. Commun. 184: 194–197, https://doi.org/10.1016/0006-291x(92)91177-r.Search in Google Scholar

Krasowska, D., Iraci, N., Santi, C., Drabowicz, J., Cieslak, M., Kaźmierczak-Barańska, J., Palomba, M., Królewska-Golińska, K., Magiera, J., and Sancineto, L. (2019). Diselenides and benzisoselenazolones as antiproliferative agents and glutathione-s-transferase inhibitors. Molecules 24: 2914, https://doi.org/10.3390/molecules24162914.Search in Google Scholar PubMed PubMed Central

Kumar, S., Sharma, N., Maurya, I.K., Bhasin, A., Wangoo, N., Brandão, P., Félix, V., Bhasin, K.K., and Sharma, R.K. (2016). Facile synthesis, structural evaluation, antimicrobial activity and synergistic effects of novel imidazo [1, 2-a]pyridine based organoselenium compounds. Eur. J. Med. Chem. 123: 916–924, https://doi.org/10.1016/j.ejmech.2016.07.076.Search in Google Scholar PubMed

Kumar, S., Johansson, H., Kanda, T., Engman, L., Müller, T., Jonsson, M., Pedulli, G.F., Petrucci, S., and Valgimigli, L. (2008). Catalytic chain-breaking pyridinol antioxidants. Org. Lett. 10: 4895–4898, https://doi.org/10.1021/ol801983n.Search in Google Scholar PubMed

Kunwar, A., Verma, P., Bhilwade, H.N., Iwaoka, M., and Priyadarsini, K.I. (2016). Dihydroxyselenolane (DHS) supplementation improves survival following whole-body irradiation (WBI) by suppressing tissue-specific inflammatory responses. Mutat. Res. Genet. Toxicol. Environ. Mutagen. 807: 33–46, https://doi.org/10.1016/j.mrgentox.2016.07.002.Search in Google Scholar PubMed

Livak, K.J. and Schmittgen, T.D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C (T)) method. Methods 25: 402–408, https://doi.org/10.1006/meth.2001.1262.Search in Google Scholar PubMed

Lo Bello, M., Oakley, A.J., Battistoni, A., Mazzetti, A.P., Nuccetelli, M., Mazzarese, G., Rossjohn, J., Parker, M.W., and Ricci, G. (1997). Multifunctional role of Tyr108 in the catalytic mechanism of human glutathione transferase P1-1. Crystallographic and kinetic studies on the Y108F Mutant Enzyme. Biochemistry 36: 6207–6217, https://doi.org/10.1021/bi962813z.Search in Google Scholar

Lu, S.C. (2009). Regulation of glutathione synthesis. Mol. Aspect. Med. 30: 42–59, https://doi.org/10.1016/j.mam.2008.05.005.Search in Google Scholar

Lushchak, V.I. (2012). Glutathione homeostasis and functions: potential targets for medical interventions. J. Amino Acids 2012: 736837.10.1155/2012/736837Search in Google Scholar

Menon, V., Thomas, R., Ghale, A.R., Reinhard, C., and Pruszak, J. (2014). Flow cytometry protocols for surface and intracellular antigen analyses of neural cell types. JoVE 94: 52241.10.3791/52241Search in Google Scholar

Parra, M., Stahl, S., and Hellmann, H. (2018). Vitamin B6 and its role in cell metabolism and physiology. Cells 7: 84, https://doi.org/10.3390/cells7070084.Search in Google Scholar

Parsanathan, R. and Jain, S.K. (2018). Hydrogen sulfide increases glutathione biosynthesis, and glucose uptake and utilisation in C2C12 mouse myotubes. Free Radic. Res. 52: 288–303, https://doi.org/10.1080/10715762.2018.1431626.Search in Google Scholar

Phadnis, P., Kunwar, A., Kumar, M., Mishra, R., Wadawale, A., Priyadarsini, K.I., and Jain, V.K. (2017). Study of polymorphism in 2, 2′-diselenobis(3-pyridinol). J. Organomet. Chem. 852: 1–7, https://doi.org/10.1016/j.jorganchem.2017.09.029.Search in Google Scholar

Pinto, M.C., Mata, A.M., and Lopez-barea, J. (1984). Reversible inactivation of Saccharomyces cerevisiae glutathione reductase under reducing conditions. Arch. Biochem. Biophys. 228: 1–12, https://doi.org/10.1016/0003-9861(84)90040-7.Search in Google Scholar

Prabhu, P., Singh, B.G., Noguchi, M., Phadnis, P.P., Jain, V.K., Iwaoka, M., and Priyadarsini, K.I. (2014). Stable selones in glutathione peroxidase-like catalytic cycle of selenonicotinamide derivative. Org. Biomol. Chem. 12: 2404–2412, https://doi.org/10.1039/c3ob42336k.Search in Google Scholar PubMed

Raghuraman, M., Verma, P., Kunwar, A., Phadnis, P.P., Jain, V.K., and Priyadarsini, K.I. (2017). Cellular evaluation of diselenonicotinamide (DSNA) as a radioprotector against cell death and DNA damage. Metallomics 9: 715–725, https://doi.org/10.1039/c7mt00034k.Search in Google Scholar PubMed

Reulecke, I., Lange, G., Albrecht, J., Klein, R., and Rarey, M. (2008). Towards an integrated description of hydrogen bonding and dehydration: decreasing false positives in virtual screening with the hyde scoring function. ChemMedChem 3: 885–897, https://doi.org/10.1002/cmdc.200700319.Search in Google Scholar

Riccardi, C. and Nicoletti, I. (2006). Analysis of apoptosis by propidium iodide staining and flow cytometry. Nat. Protoc. 1: 1458–1461, https://doi.org/10.1038/nprot.2006.238.Search in Google Scholar

Sausen de Freitas, A., de Souza Prestes, A., Wagner, C., HaigertSudati, J., Alves, D., Oliveira Porciúncula, L., Kade, I.J., and Teixeira Rocha, J.B. (2010). Reduction of diphenyl diselenide and analogs by mammalian thioredoxin reductase is independent of their glutathione peroxidase-like activity: a possible novel pathway for their antioxidant activity. Molecules 15: 7699–7714, https://doi.org/10.3390/molecules15117700.Search in Google Scholar

Schneider, N., Hindle, S., Lange, G., Klein, R., Albrecht, J., Briem, H., Beyer, K., Claußen, H., Gastreich, M., Lemmen, C., et al.. (2012). Substantial improvements in large-scale redocking and screening using the novel HYDE scoring function. J. Comput. Aided Mol. Des. 26: 701–723, https://doi.org/10.1007/s10822-011-9531-0.Search in Google Scholar

Schneider, N., Lange, G., Hindle, S., Klein, R., and Rarey, M. (2013). A consistent description of hydrogen bond and dehydration energies in protein–ligand complexes: methods behind the HYDE scoring function. J. Comput. Aided Mol. Des. 27: 15–29, https://doi.org/10.1007/s10822-012-9626-2.Search in Google Scholar

Schöning-Stierand, K., Diedrich, K., Fährrolfes, R., Flachsenberg, F., Meyder, A., Nittinger, E., Steinegger, R., and Rarey, M. (2020). ProteinsPlus: interactive analysis of protein–ligand binding interfaces. Nucleic Acids Res. 48: W48–W53, https://doi.org/10.1093/nar/gkaa235.Search in Google Scholar

Singh, V.P., Poon, J.F., Butcher, R.J., and Engman, L. (2014). Pyridoxine-derived organoselenium compounds with glutathione peroxidase-like and chain-breaking antioxidant activity. Chemistry 20: 12563–12571, https://doi.org/10.1002/chem.201403229.Search in Google Scholar

Sudati, J.H., Nogara, P.A., Saraiva, R.A., Wagner, C., Alberto, E.E., Braga, A.L., Fachinetto, R., Piquini, P.C., and Rocha, J.B.T. (2018). Diselenoamino acid derivatives as GPx mimics and as substrates of TrxR: in vitro and in silico studies. Org. Biomol. Chem. 16: 3777–3787, https://doi.org/10.1039/c8ob00451j.Search in Google Scholar

Tan, H.W., Mo, H.Y., Lau, A., and Xu, Y.M. (2018). Selenium species: current status and potentials in cancer prevention and therapy. Int. J. Mol. Sci. 20: 75, https://doi.org/10.3390/ijms20010075.Search in Google Scholar

Tappel, A.L. (1978). Glutathione peroxidase and hydroperoxydes. Methods Enzymol. 52: 506–513.10.1016/S0076-6879(78)52055-7Search in Google Scholar

Volkamer, A., Griewel, A., Grombacher, T., and Rarey, M. (2010). Analyzing the topology of active sites: on the prediction of pockets and subpockets. J. Chem. Inf. Model. 50: 2041–2052, https://doi.org/10.1021/ci100241y.Search in Google Scholar PubMed

Yoo, M.H., Xu, X.M., Carlson, B.A., Patterson, A.D., Gladyshev, V.N., and Hatfield, D.L. (2007). Targeting thioredoxin reductase 1 reduction in cancer cells inhibits self-sufficient growth and DNA replication. PLoS One 2: e1112, https://doi.org/10.1371/journal.pone.0001112.Search in Google Scholar PubMed PubMed Central

Zhang, H., Limphong, P., Pieper, J., Liu, Q., Rodesch, C.K., Christians, E., and Benjamin, I.J. (2012). Glutathione-dependent reductive stress triggers mitochondrial oxidation and cytotoxicity. Faseb. J. 26: 1442–1451, https://doi.org/10.1096/fj.11-199869.Search in Google Scholar PubMed PubMed Central

Zhang, L. and Tew, K.D. (2021). Reductive stress in cancer. Adv. Cancer Res. 152: 383–413.10.1016/bs.acr.2021.03.009Search in Google Scholar PubMed

Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/hsz-2022-0123).

Received: 2022-02-18
Accepted: 2022-08-01
Published Online: 2022-08-25
Published in Print: 2022-09-27

© 2022 Walter de Gruyter GmbH, Berlin/Boston

Articles in the same Issue

  1. Frontmatter
  2. Research Articles/Short Communications
  3. Cell Biology and Signaling
  4. Diselenide-derivative of 3-pyridinol targets redox enzymes leading to cell cycle deregulation and apoptosis in A549 cells
  5. NF90/NFAR (nuclear factors associated with dsRNA) – a new methylation substrate of the PRMT5-WD45-RioK1 complex
  6. PHF20L1 mediates PAX2 expression to promote angiogenesis and liver metastasis in colorectal cancer through regulating HIC1
  7. Nitazoxanide inhibits osteosarcoma cells growth and metastasis by suppressing AKT/mTOR and Wnt/β-catenin signaling pathways
  8. LncRNA-p21 suppresses cell proliferation and induces apoptosis in gastric cancer by sponging miR-514b-3p and up-regulating ARHGEF9 expression
  9. L-theanine induces skeletal muscle fiber type transformation by activation of prox1/CaN signaling pathway in C2C12 myotubes
  10. Proteolysis
  11. TMPRSS13 zymogen activation, surface localization, and shedding is regulated by proteolytic cleavage within the non-catalytic stem region
https://doi.org/10.1515/hsz-2022-0123
Keywords for this article
apoptosis; cell cycle arrest; organodiselenides; reactive oxygen species; redox modulation

Articles in the same Issue

  1. Frontmatter
  2. Research Articles/Short Communications
  3. Cell Biology and Signaling
  4. Diselenide-derivative of 3-pyridinol targets redox enzymes leading to cell cycle deregulation and apoptosis in A549 cells
  5. NF90/NFAR (nuclear factors associated with dsRNA) – a new methylation substrate of the PRMT5-WD45-RioK1 complex
  6. PHF20L1 mediates PAX2 expression to promote angiogenesis and liver metastasis in colorectal cancer through regulating HIC1
  7. Nitazoxanide inhibits osteosarcoma cells growth and metastasis by suppressing AKT/mTOR and Wnt/β-catenin signaling pathways
  8. LncRNA-p21 suppresses cell proliferation and induces apoptosis in gastric cancer by sponging miR-514b-3p and up-regulating ARHGEF9 expression
  9. L-theanine induces skeletal muscle fiber type transformation by activation of prox1/CaN signaling pathway in C2C12 myotubes
  10. Proteolysis
  11. TMPRSS13 zymogen activation, surface localization, and shedding is regulated by proteolytic cleavage within the non-catalytic stem region
Sign up now to receive a 20% welcome discount
Institutional Access
How does access work?
Have an idea on how to improve our website?
Please write us.
© 2026 De Gruyter Brill
Downloaded on 30.1.2026 from https://www.degruyterbrill.com/document/doi/10.1515/hsz-2022-0123/html
Scroll to top button