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Usnic Acid Exerts Antiproliferative and Apoptotic Effects by Suppressing NF-B p50 in DU145 Cells

Year 2023, Volume: 82 Issue: 2, 251 - 257, 21.12.2023
https://doi.org/10.26650/EurJBiol.2023.1274707

Abstract

Objective: Nuclear factor kappa B (NF-κB) is one pathway that controls the expression of genes involved in many cancer events such as proliferation, apoptosis, metastasis, and invasion. Usnic acid is a molecule with many biological effects such as being anticholinergic, gastroprotective, anti-inflammatory, anti-cancerous, and especially antioxidant. This study aims to mechanistically examine the apoptotic behaviors of usnic acid in DU145 prostate cancer cells and the molecules it acts on in the NF-κB pathway.

Materials and Methods: This study investigates the apoptotic changes in DU145 cells after usnic acid administration through JC-1 staining and caspase-3 activity measurements. In addition, it tests the effects of usnic acid on subunit p50 and p65 protein and gene expressions in the NF-κB pathway through the respective Western blot and qPCR measurements.

Results: The IC50 values of usnic acid at 24 and 48 h in DU145 cells were calculated as 167.06±12.35 μM and 42.15±3.76 μM, respectively. In addition, JC-1 staining showed usnic acid-treated DU145 cells to trigger apoptosis by increasing the membrane permeability of their mitochondria. NF-κB p50 protein expression was also found to be suppressed after usnic acid administration.

Conclusion: The results of this study show usnic acid administration to suppress proliferation and to induce mitochondrial apoptosis by suppressing the NF-κB pathway in DU145 cells. This effect of usnic acid indicates it to be combinable with chemotherapeutic agents and evaluable as an alternative in cancer treatment.

References

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  • Kohlhardt-Floehr C, Boehm F, Troppens S, Lademann J, Truscott TG. Prooxidant and antioxidant behaviour of usnic acid from lichens under UVB-light irradiation-Studies on human cells. J Photochem Photobiol B. 2010;101(1):97-102. google scholar
  • Han D, Matsumaru K, Rettori D, Kaplowitz N. Usnic acid-induced necrosis of cultured mouse hepatocytes: Inhibition of mitochondrial function and oxidative stress. Biochem Pharma-col. 2004;67(3):439-451. google scholar
  • Pramyothin P, Janthasoot W, Pongnimitprasert N, Phrukudom S, Ruangrungsi N. Hepatotoxic effect of (+) usnic acid from Usnea siamensis Wainio in rats, isolated rat hepatocytes and isolated rat liver mitochondria. J Ethnopharmacol. 2004;90(2-3): 381-387. google scholar
  • Sonko BJ, Schmitt TC, Guo L, et al. Assessment of usnic acid toxicity in rat primary hepatocytes using 13C isotopomer dis-tribution analysis of lactate, glutamate and glucose. Food Chem Toxicol. 2011;49(11):2968-2974. google scholar
  • Sahu SC, Amankwa-Sakyi M, O’Donnell Jr MW, Sprando RL. Effects of usnic acid exposure on human hepatoblastoma HepG2 cells in culture. J Appl Toxicol. 2012;32(9):722-730. google scholar
  • Mariappan N, Elks CM, Sriramula S, et al. NF-kB-induced oxida-tive stress contributes to mitochondrial and cardiac dysfunction in type II diabetes. Cardiovasc Res. 2010;85(3):473-483. google scholar
  • Elks CM, Mariappan N, Haque M, Guggilam A, Majid DS, Fran-cis J. Chronic NF-kB blockade reduces cytosolic and mitochon-drial oxidative stress and attenuates renal injury and hypertension in SHR. Am J Physiol Renal Physiol. 2009;296(2):298-305. google scholar
  • Takai M, Uehara Y, Beisler JA. Usnic acid derivatives as potential antineoplastic agents. JMed Chem. 1979; 22(11):1380-1384. google scholar
  • Cardarelli M, Serino G, Campanella L, et al. Antimitotic effects of usnic acid on different biological systems. Cell Mol Life Sci. 1997;53:667-672. google scholar
  • Backorova M, Jendzelovsky R, Kello M, Backor M, Mikes J, Fedorocko P. Lichen secondary metabolites are responsible for induction of apoptosis in HT-29 and A2780 human cancer cell lines. Toxicol In Vitro. 2012;26(3): 462-468. google scholar
  • Dincsoy AB, Duman DC. Changes in apoptosis-related gene ex-pression profiles in cancer cell lines exposed to usnic acid lichen secondary metabolite. Turk J Biol. 2017;41(3): 484-493. google scholar
  • Jin JQ, Li CQ, He LC. Down-regulatory effect of usnic acid on nu-clear factor-KB-dependent tumor necrosis factor-a and inducible nitric oxide synthase expression in lipopolysaccharide-stimulated macrophages RAW 264.7. Phytother Res. 2008; 22(12):1605-1609. google scholar
Year 2023, Volume: 82 Issue: 2, 251 - 257, 21.12.2023
https://doi.org/10.26650/EurJBiol.2023.1274707

Abstract

References

  • Sung H, Ferlay J, Siegel RL, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: Cancer J Clin. 2021;71(3):209-249. google scholar
  • Gandaglia G, Leni R, Bray F, et al. Epidemiology and prevention of prostate cancer. Eur Urol Oncol. 2021;4(6):877-892. google scholar
  • Rawla P. Epidemiology of prostate cancer. World J Oncol. 2019;10(2):63-89. google scholar
  • Hurwitz M. Chemotherapy in prostate cancer. Curr Oncol Rep. 2015;17:1-10. google scholar
  • Erstad DJ, Cusack JC. Targeting the NF-kB pathway in cancer therapy. Surg Oncol Clin. 2013;22(4):705-746. google scholar
  • Rinkenbaugh AL, Baldwin AS. The NF-kB pathway and cancer stem cells. Cells. 2016;5(2):16. doi: 10.3390/cells5020016. google scholar
  • Ingolfsdottir K. Usnic acid. Phytochem.2002;61(7):729-736. google scholar
  • Araujo AA, de Melo MG, Rabelo TK, et al. Review of the bi-ological properties and toxicity of usnic acid. Nat Prod Res. 2015;29(23):2167-2180. google scholar
  • Cakmak KC, Gulcin I. Anticholinergic and antioxidant activ-ities of usnic acid-an activity-structure insight. Toxicol Rep. 2019;6:1273-1280. google scholar
  • Hoa NT, Van Bay M, Mechler A, Vo QV. Is usnic acid a promising radical scavenger? ACS Omega. 2020;5(28):17715-17720. google scholar
  • Maulidiyah M, Rachman F, Mulkiyan OMZ, et al. Antioxidant activity of usnic acid compound from methanol extract of Lichen usnea sp. J Oleo Sci. 2023;72(2):179-188. google scholar
  • Erdoğan Ö, Paşa S, Demirbolat GM, Çevik Ö. Green biosynthesis, characterization, and cytotoxic effect of magnetic iron nanopar-ticles using Brassica oleracea var capitata sub var rubra (red cabbage) aqueous peel extract. Turk J Chem. 2021; 45(4):1086-1096. google scholar
  • Sivandzade F, Bhalerao A, Cucullo L. Analysis of the mito-chondrial membrane potential using the cationic JC-1 dye as a sensitive fluorescent probe. Bio-protoc. 2019;9(1):e3128. doi: 10.21769/BioProtoc.3128. google scholar
  • Erdogan O, Cevik O. Myricetin can control metastasis and invasion by suppressing ATF2-related signaling pathway in Rapamycin-resistant HepG2 hepatocellular cancer cells. J Res Pharm. 2023;27(2):557-565. google scholar
  • Pasa S, Erdogan O, Cevik O. Design, synthesis and investiga-tion of procaine based new Pd complexes as DNA methyltrans-ferase inhibitor on gastric cancer cells. Inorg Chem Commun. 2021;132:108846. https://doi.org/10.1016/j.inoche.2021.108846. google scholar
  • Inoue Ji, Gohda J, Akiyama T, Semba K. NF-B activation in devel-opment and progression of cancer. Cancer Sci. 2007;98(3):268-274. google scholar
  • Sun SC. The non-canonical NF-B pathway in immunity and in-flammation. Nat Rev Immunol. 2017;17(9):545-558. google scholar
  • Naugler WE, Karin M. NF-B and cancer—identifying targets and mechanisms. Curr Opin Genet Dev. 2008;18(1):19-26. google scholar
  • Yamamoto Y, Gaynor RB. Therapeutic potential of inhibition of the NF-kB pathway in the treatment of inflammation and cancer. J Clin Invest. 2001;107(2):135-142. google scholar
  • Ramadass V, Vaiyapuri T, Tergaonkar V. Small molecule NF-kB pathway inhibitors in clinic. Int J Mol Sci. 2020;21(14):5164. doi: 10.3390/ijms21145164 google scholar
  • Cevik O, Acidereli H, Turut FA, Yildirim S, Acilan C. Cabazitaxel exhibits more favorable molecular changes compared to other taxanes in androgen-independent prostate cancer cells. J Biochem Mol Toxicol. 2020;34(9):e22542. doi: 10.1002/jbt.22542. google scholar
  • Odabasoglu F, Cakir A, Suleyman H, et al. Gastroprotective and antioxidant effects of usnic acid on indomethacin-induced gastric ulcer in rats. J Ethnopharmacol. 2006; 103(1):59-65. google scholar
  • Kohlhardt-Floehr C, Boehm F, Troppens S, Lademann J, Truscott TG. Prooxidant and antioxidant behaviour of usnic acid from lichens under UVB-light irradiation-Studies on human cells. J Photochem Photobiol B. 2010;101(1):97-102. google scholar
  • Han D, Matsumaru K, Rettori D, Kaplowitz N. Usnic acid-induced necrosis of cultured mouse hepatocytes: Inhibition of mitochondrial function and oxidative stress. Biochem Pharma-col. 2004;67(3):439-451. google scholar
  • Pramyothin P, Janthasoot W, Pongnimitprasert N, Phrukudom S, Ruangrungsi N. Hepatotoxic effect of (+) usnic acid from Usnea siamensis Wainio in rats, isolated rat hepatocytes and isolated rat liver mitochondria. J Ethnopharmacol. 2004;90(2-3): 381-387. google scholar
  • Sonko BJ, Schmitt TC, Guo L, et al. Assessment of usnic acid toxicity in rat primary hepatocytes using 13C isotopomer dis-tribution analysis of lactate, glutamate and glucose. Food Chem Toxicol. 2011;49(11):2968-2974. google scholar
  • Sahu SC, Amankwa-Sakyi M, O’Donnell Jr MW, Sprando RL. Effects of usnic acid exposure on human hepatoblastoma HepG2 cells in culture. J Appl Toxicol. 2012;32(9):722-730. google scholar
  • Mariappan N, Elks CM, Sriramula S, et al. NF-kB-induced oxida-tive stress contributes to mitochondrial and cardiac dysfunction in type II diabetes. Cardiovasc Res. 2010;85(3):473-483. google scholar
  • Elks CM, Mariappan N, Haque M, Guggilam A, Majid DS, Fran-cis J. Chronic NF-kB blockade reduces cytosolic and mitochon-drial oxidative stress and attenuates renal injury and hypertension in SHR. Am J Physiol Renal Physiol. 2009;296(2):298-305. google scholar
  • Takai M, Uehara Y, Beisler JA. Usnic acid derivatives as potential antineoplastic agents. JMed Chem. 1979; 22(11):1380-1384. google scholar
  • Cardarelli M, Serino G, Campanella L, et al. Antimitotic effects of usnic acid on different biological systems. Cell Mol Life Sci. 1997;53:667-672. google scholar
  • Backorova M, Jendzelovsky R, Kello M, Backor M, Mikes J, Fedorocko P. Lichen secondary metabolites are responsible for induction of apoptosis in HT-29 and A2780 human cancer cell lines. Toxicol In Vitro. 2012;26(3): 462-468. google scholar
  • Dincsoy AB, Duman DC. Changes in apoptosis-related gene ex-pression profiles in cancer cell lines exposed to usnic acid lichen secondary metabolite. Turk J Biol. 2017;41(3): 484-493. google scholar
  • Jin JQ, Li CQ, He LC. Down-regulatory effect of usnic acid on nu-clear factor-KB-dependent tumor necrosis factor-a and inducible nitric oxide synthase expression in lipopolysaccharide-stimulated macrophages RAW 264.7. Phytother Res. 2008; 22(12):1605-1609. google scholar
There are 34 citations in total.

Details

Primary Language English
Subjects Plant Cell and Molecular Biology, Animal Cell and Molecular Biology
Journal Section Themed Articles - Research Articles
Authors

Ömer Erdoğan 0000-0002-8327-7077

Burçin İrem Abas 0000-0002-1018-5577

Özge Çevik 0000-0002-9325-3757

Publication Date December 21, 2023
Submission Date March 31, 2023
Published in Issue Year 2023 Volume: 82 Issue: 2

Cite

AMA Erdoğan Ö, Abas Bİ, Çevik Ö. Usnic Acid Exerts Antiproliferative and Apoptotic Effects by Suppressing NF-B p50 in DU145 Cells. Eur J Biol. December 2023;82(2):251-257. doi:10.26650/EurJBiol.2023.1274707