Araştırma Makalesi
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Anticancer Effect of Usnea longissima Ach. Ethanolic Extract in HCT116 Colon Cancer Cells and Biofilm Inhibition Capacity

Yıl 2025, Cilt: 15 Sayı: 3, 409 - 416, 24.09.2025
https://doi.org/10.33631/sabd.1666913

Öz

Aim: Usnea longissima Ach. has been used historically in herbal medicine for its antibiotic properties. However, its possible anticancer effects are still being investigated.
Material and Methods: HCT116 colon cancer and HUVEC cell lines were treated with U. longissima Ach. ethanolic extract at 25-800 µg/mL concentrations and cell viability analysed with the WST-1 assay. Human Caspase-3, -8 and -9 ELISA kits were used to investigate the apoptotic effects of extract in HCT116 cells. Migration effects were evaluated on HCT116 cells by wound healing assay. Biofilm inhibition was evaluated using the microplate method, and anti-quorum sensing (QS) effect was determined the well diffusion method.
Results: Ethanolic extract of U. longissima Ach. reduced the cell viability level to 14% in HCT116 colon cancer cells at 800 µg/mL. In HUVEC cells, the non-significant decrease in cell viability indicated that the U. longissima Ach. had minimal cytotoxic effects on healthy cells. Caspase-3 and Caspase-9 expression levels in HCT116 cells increased approximately 5-fold compared to the control group, indicating that the extract induced intrinsic apoptotic response. Additionally, the extract significantly inhibited the HCT116 cell migration level. In addition to the anti-QS effect of U. longissima Ach. ethanolic extract (opaque zone 10-12 mm), it also reduced the biofilm formation of Escherichia coli bacteria, which is dense in the microbiota, by 80-93%.
Conclusion: In conclusion, it is thought that the significant anticancer and antibiofilm effects of U. longissima Ach. ethanolic extract on colon cancer cells may be promising as a complement to conventional chemotherapeutics after being confirmed by detailed studies at the in vivo level.

Kaynakça

  • Molins A, Moya PO, Garcia-Breijo FJ, José RA, Barreno E. A multi-tool approach to assess microalgal diversity in lichens: isolation, Sanger sequencing, HTS and ultrastructural correlations. Lichenologist. 2018;50(1):123-38.
  • Nayanakantha NMC, Gajameragedara S. A survey of lichens in the Kandy Municipal Region. Univ Peradeniya. 2003;31:35-41.
  • Halici M, Odabasoglu F, Suleyman H, Cakir A, Aslan A, Bayir Y. Effects of water extract of Usnea longissima on antioxidant enzyme activity and mucosal damage caused by indomethacin in rats. Phytomedicine. 2005;12(9):656-62.
  • Reddy SD, Siva B, Kumar K, Babu VP, Sravanthi V, Boustie J, et al. Comprehensive analysis of secondary metabolites in Usnea longissima (Lichenized Ascomycetes, Parmeliaceae) using UPLC-ESI-QTOF-MS/MS and pro-apoptotic activity of barbatic acid. Molecules. 2019;24(12):2270.
  • Chandra M. Antimicrobial activity of medicinal plants against human pathogenic bacteria. Int J Biotechnol Bioeng Res. 2013;4(7):653-58.
  • Aiello P, Sharghi M, Mansourkhani SM, Ardekan AP, Jouybari L, Daraei N, et al. Medicinal plants in the prevention and treatment of colon cancer. Oxid Med Cell Longev. 2019;2019(1):2075614.
  • Dejea CM, Sears CL. Do biofilms confer a pro-carcinogenic state? Gut Microbes. 2016;7(1):54-7.
  • Tomkovich S, Dejea CM, Winglee K, Drewes JL, Chung L, Housseau F, et al. Human colon mucosal biofilms from healthy or colon cancer hosts are carcinogenic. J Clin Invest. 2019;129(4):1699-712.
  • Chew SS, Tan LTH, Law JWF, Pusparajah P, Goh BH, Ab Mutalib NS, et al. Targeting gut microbial biofilms—a key to hinder colon carcinogenesis? Cancers. 2020;12(8):2272.
  • Sionov RV, Steinberg D. Targeting the holy triangle of quorum sensing, biofilm formation, and antibiotic resistance in pathogenic bacteria. Microorganisms. 2022;10(6):1239.
  • Esmeeta A, Adhikary S, Dharshnaa V, Swarnamughi P, Maqsummiya ZU, Banerjee A, et al. Plant-derived bioactive compounds in colon cancer treatment: an updated review. Biomed Pharmacother. 2022;153:113384.
  • Popovici V, Bucur L, Vochita G, Gherghel D, Mihai CT, Rambu D, et al. In vitro anticancer activity and oxidative stress biomarkers status determined by Usnea barbata (L.) FH Wigg. dry extracts. Antioxidants. 2021;10(7):1141.
  • Tuong TL, Do LT, Aree T, Wonganan P, Chavasiri W. Tetrahydroxanthone–chromanone heterodimers from lichen Usnea aciculifera and their cytotoxic activity against human cancer cell lines. Fitoterapia. 2020;147:104732.
  • Engin TA, Emsen B, Yılmaz R, Koc RC, İnan B, Ozcimen D. Cytotoxicity of Usnea longissima Ach. extracts and its secondary metabolite, usnic acid on different cells. Anatol J Bot. 2023;7(2):140-5.
  • Ghosh S, Basu S, Anbarasu A, Ramaiah S. A comprehensive review of antimicrobial agents against clinically important bacterial pathogens: prospects for phytochemicals. Phytother Res. 2025;39(1):138-61.
  • Bharti S, Nayaka S. Evaluation of some traditional therapeutic properties of Usnea longissima (Ascomycota, lichenized fungi): antimicrobial, antiquorum and antioxidant. J Microbiol Biotechnol Food Sci. 2022;11(4):e3163.
  • Dülger B, Dülger G, Gedik B, Oran S. Evaluation on antimicrobial activity of the lichen Pleurosticta acetabulum. Düzce Univ Bilim Teknol Derg. 2023;11(1):199-203.
  • Akkoyunlu A, Dulger G. Comparative in vitro study of antimicrobial, antibiofilm and quorum sensing inhibitory activities of Hypericum calycinum L. and Parietaria officinalis L. extracts. Sağlık Bilim Değer. 2024;14(3):363-8.
  • Stepanović S, Vuković D, Dakić I, et al. A modified microtiter-plate test for quantification of staphylococcal biofilm formation. J Microbiol Methods. 2000;40:175-9.
  • Akkoyunlu A, Dülger G. Exploring the antibiofilm effects on Escherichia coli biofilm associated with colon cancer and anticancer activities on HCT116 cell line of bee products. Biofouling. 2024;40(3-4):235-44.
  • Şahin N, Emsen B, Aslan A, Sadi G. Antioxidant potential of Pseudevernia furfuracea (L.) Zopf and its secondary metabolites on hepatocellular carcinoma cells: regulation of antioxidant enzymes. Anatol J Bot. 2021;5(2):127-33.
  • Ureña-Vacas I, González-Burgos E, Divakar PK, Gómez-Serranillos MP. Lichen depsidones with biological interest. Planta Med. 2022;88(11):855-80.
  • Dandapat M, Paul S. Secondary metabolites from lichen Usnea longissima and its pharmacological relevance. Pharmacogn Res. 2019;11(2): 103.
  • Sepahvand A, Studzińska-Sroka E, Ramak P, Karimian V. Usnea sp.: antimicrobial potential, bioactive compounds, ethnopharmacological uses and other pharmacological properties; a review article. J Ethnopharmacol. 2021;268:113656.
  • Altuner D, Ekşi S, Süleyman H, Turumtay EA. The effect of Usnea longissima extract on lung cancer cell line (A549). ABC Res. 2021;3(1):1-5.
  • Kaeser MD, Pebernard S, Iggo RD. Regulation of p53 stability and function in HCT116 colon cancer cells. J Biol Chem. 2004;279(9):7598-605.
  • Plaskova A, Mlcek J. New insights of the application of water or ethanol-water plant extract rich in active compounds in food. Front Nutr. 2023;10:1118761.
  • Pistritto G, Trisciuoglio D, Ceci C, Garufi A, D'Orazi G. Apoptosis as anticancer mechanism: function and dysfunction of its modulators and targeted therapeutic strategies. Aging (Albany NY). 2016;8(4):603.
  • Choi E, Murray B, Choi S. Biofilm and cancer: interactions and future directions for cancer therapy. Int J Mol Sci. 2023;24(16):12836.
  • Alhinai EA, Walton GE, Commane DM. The role of the gut microbiota in colorectal cancer causation. Int J Mol Sci. 2019;20(21):5295.
  • Wang Y, Bian Z, Wang Y. Biofilm formation and inhibition mediated by bacterial quorum sensing. Appl Microbiol Biotechnol. 2022;106(19):6365-81.
  • Francolini I, Norris P, Piozzi A, Donelli G, Stoodley P. Usnic acid, a natural antimicrobial agent able to inhibit bacterial biofilm formation on polymer surfaces. Antimicrob Agents Chemother. 2004;48(11):4360-5.
  • Nithyanand P, Beema Shafreen RM, Muthamil S, Karutha Pandian S. Usnic acid, a lichen secondary metabolite inhibits Group A Streptococcus biofilms. Antonie Van Leeuwenhoek. 2015;107:263-72.
  • Grumezescu AM, Cotar AI, Andronescu E, Ficai A, Ghitulica CD, Grumezescu V, et al. In vitro activity of the new water-dispersible Fe₃O₄@usnic acid nanostructure against planktonic and sessile bacterial cells. J Nanopart Res. 2013;15:1766.

Usnea longissima Ach. Etanolik Özütünün HCT116 Kolon Kanseri Hücrelerinde Antikanser Etkisi ve Biyofilm İnhibisyon Kapasitesi

Yıl 2025, Cilt: 15 Sayı: 3, 409 - 416, 24.09.2025
https://doi.org/10.33631/sabd.1666913

Öz

Amaç: Usnea longissima Ach. antibiyotik özellikleri nedeniyle geleneksel olarak bitkisel tedavide kullanılmıştır. Ancak, olası antikanser etkileri halen araştırılmaktadır.
Gereç ve Yöntemler: HCT116 kolon kanseri ve HUVEC hücre hatları, 25-800 µg/mL konsantrasyonlarda U. longissima Ach. etanolik özütü ile muamele edildi ve hücre canlılığı WST-1 testi ile analiz edildi. İnsan Kaspaz-3, -8 ve -9 ELISA kitleri, özütün HCT116 hücrelerindeki apoptotik etkilerini araştırmak için kullanıldı. Migrasyon etkileri, HCT116 hücrelerinde yara iyileştirme testi ile değerlendirildi. Biyofilm inhibisyonu mikroplate yöntemi ile değerlendirildi ve anti-quorum sensing (QS) etkisi kuyu difüzyon yöntemi ile belirlendi.
Bulgular: U. longissima Ach. etanolik özütü, 800 µg/mL'de HCT116 kolon kanseri hücrelerinde hücre canlılığı seviyesini %14'e düşürdü. HUVEC hücrelerinde, hücre canlılığındaki anlamlı olmayan azalma, U. longissima Ach.'nın sağlıklı hücreler üzerinde minimal sitotoksik etkiye sahip olduğunu gösterdi. HCT116 hücrelerinde Kaspaz-3 ve Kaspaz-9 ekspresyon seviyeleri kontrol grubuna kıyasla yaklaşık 5 kat arttı ve bu da özütün içsel apoptotik yanıtı indüklediğini gösterdi. Ek olarak, özüt HCT116 hücre migrasyon seviyesini önemli ölçüde engelledi. U. longissima Ach. etanol özütü anti-QS etkisine (opak bölge 10-12 mm) ek olarak, mikrobiyota da yoğun olan Escherichia coli bakterisinin biyofilm oluşumunu %80-93 oranında azalttı.
Sonuç: Sonuç olarak, U. longissima Ach. etanol ekstresinin kolon kanseri hücreleri üzerindeki önemli antikanser ve antibiyofilm etkilerinin, in vivo düzeyde yapılan detaylı çalışmalarla doğrulanmasının ardından konvansiyonel kemoterapötiklere tamamlayıcı olarak umut verici olabileceği düşünülmektedir.

Etik Beyan

In vitro bir çalışma olduğundan etik kurula gerek yoktur

Kaynakça

  • Molins A, Moya PO, Garcia-Breijo FJ, José RA, Barreno E. A multi-tool approach to assess microalgal diversity in lichens: isolation, Sanger sequencing, HTS and ultrastructural correlations. Lichenologist. 2018;50(1):123-38.
  • Nayanakantha NMC, Gajameragedara S. A survey of lichens in the Kandy Municipal Region. Univ Peradeniya. 2003;31:35-41.
  • Halici M, Odabasoglu F, Suleyman H, Cakir A, Aslan A, Bayir Y. Effects of water extract of Usnea longissima on antioxidant enzyme activity and mucosal damage caused by indomethacin in rats. Phytomedicine. 2005;12(9):656-62.
  • Reddy SD, Siva B, Kumar K, Babu VP, Sravanthi V, Boustie J, et al. Comprehensive analysis of secondary metabolites in Usnea longissima (Lichenized Ascomycetes, Parmeliaceae) using UPLC-ESI-QTOF-MS/MS and pro-apoptotic activity of barbatic acid. Molecules. 2019;24(12):2270.
  • Chandra M. Antimicrobial activity of medicinal plants against human pathogenic bacteria. Int J Biotechnol Bioeng Res. 2013;4(7):653-58.
  • Aiello P, Sharghi M, Mansourkhani SM, Ardekan AP, Jouybari L, Daraei N, et al. Medicinal plants in the prevention and treatment of colon cancer. Oxid Med Cell Longev. 2019;2019(1):2075614.
  • Dejea CM, Sears CL. Do biofilms confer a pro-carcinogenic state? Gut Microbes. 2016;7(1):54-7.
  • Tomkovich S, Dejea CM, Winglee K, Drewes JL, Chung L, Housseau F, et al. Human colon mucosal biofilms from healthy or colon cancer hosts are carcinogenic. J Clin Invest. 2019;129(4):1699-712.
  • Chew SS, Tan LTH, Law JWF, Pusparajah P, Goh BH, Ab Mutalib NS, et al. Targeting gut microbial biofilms—a key to hinder colon carcinogenesis? Cancers. 2020;12(8):2272.
  • Sionov RV, Steinberg D. Targeting the holy triangle of quorum sensing, biofilm formation, and antibiotic resistance in pathogenic bacteria. Microorganisms. 2022;10(6):1239.
  • Esmeeta A, Adhikary S, Dharshnaa V, Swarnamughi P, Maqsummiya ZU, Banerjee A, et al. Plant-derived bioactive compounds in colon cancer treatment: an updated review. Biomed Pharmacother. 2022;153:113384.
  • Popovici V, Bucur L, Vochita G, Gherghel D, Mihai CT, Rambu D, et al. In vitro anticancer activity and oxidative stress biomarkers status determined by Usnea barbata (L.) FH Wigg. dry extracts. Antioxidants. 2021;10(7):1141.
  • Tuong TL, Do LT, Aree T, Wonganan P, Chavasiri W. Tetrahydroxanthone–chromanone heterodimers from lichen Usnea aciculifera and their cytotoxic activity against human cancer cell lines. Fitoterapia. 2020;147:104732.
  • Engin TA, Emsen B, Yılmaz R, Koc RC, İnan B, Ozcimen D. Cytotoxicity of Usnea longissima Ach. extracts and its secondary metabolite, usnic acid on different cells. Anatol J Bot. 2023;7(2):140-5.
  • Ghosh S, Basu S, Anbarasu A, Ramaiah S. A comprehensive review of antimicrobial agents against clinically important bacterial pathogens: prospects for phytochemicals. Phytother Res. 2025;39(1):138-61.
  • Bharti S, Nayaka S. Evaluation of some traditional therapeutic properties of Usnea longissima (Ascomycota, lichenized fungi): antimicrobial, antiquorum and antioxidant. J Microbiol Biotechnol Food Sci. 2022;11(4):e3163.
  • Dülger B, Dülger G, Gedik B, Oran S. Evaluation on antimicrobial activity of the lichen Pleurosticta acetabulum. Düzce Univ Bilim Teknol Derg. 2023;11(1):199-203.
  • Akkoyunlu A, Dulger G. Comparative in vitro study of antimicrobial, antibiofilm and quorum sensing inhibitory activities of Hypericum calycinum L. and Parietaria officinalis L. extracts. Sağlık Bilim Değer. 2024;14(3):363-8.
  • Stepanović S, Vuković D, Dakić I, et al. A modified microtiter-plate test for quantification of staphylococcal biofilm formation. J Microbiol Methods. 2000;40:175-9.
  • Akkoyunlu A, Dülger G. Exploring the antibiofilm effects on Escherichia coli biofilm associated with colon cancer and anticancer activities on HCT116 cell line of bee products. Biofouling. 2024;40(3-4):235-44.
  • Şahin N, Emsen B, Aslan A, Sadi G. Antioxidant potential of Pseudevernia furfuracea (L.) Zopf and its secondary metabolites on hepatocellular carcinoma cells: regulation of antioxidant enzymes. Anatol J Bot. 2021;5(2):127-33.
  • Ureña-Vacas I, González-Burgos E, Divakar PK, Gómez-Serranillos MP. Lichen depsidones with biological interest. Planta Med. 2022;88(11):855-80.
  • Dandapat M, Paul S. Secondary metabolites from lichen Usnea longissima and its pharmacological relevance. Pharmacogn Res. 2019;11(2): 103.
  • Sepahvand A, Studzińska-Sroka E, Ramak P, Karimian V. Usnea sp.: antimicrobial potential, bioactive compounds, ethnopharmacological uses and other pharmacological properties; a review article. J Ethnopharmacol. 2021;268:113656.
  • Altuner D, Ekşi S, Süleyman H, Turumtay EA. The effect of Usnea longissima extract on lung cancer cell line (A549). ABC Res. 2021;3(1):1-5.
  • Kaeser MD, Pebernard S, Iggo RD. Regulation of p53 stability and function in HCT116 colon cancer cells. J Biol Chem. 2004;279(9):7598-605.
  • Plaskova A, Mlcek J. New insights of the application of water or ethanol-water plant extract rich in active compounds in food. Front Nutr. 2023;10:1118761.
  • Pistritto G, Trisciuoglio D, Ceci C, Garufi A, D'Orazi G. Apoptosis as anticancer mechanism: function and dysfunction of its modulators and targeted therapeutic strategies. Aging (Albany NY). 2016;8(4):603.
  • Choi E, Murray B, Choi S. Biofilm and cancer: interactions and future directions for cancer therapy. Int J Mol Sci. 2023;24(16):12836.
  • Alhinai EA, Walton GE, Commane DM. The role of the gut microbiota in colorectal cancer causation. Int J Mol Sci. 2019;20(21):5295.
  • Wang Y, Bian Z, Wang Y. Biofilm formation and inhibition mediated by bacterial quorum sensing. Appl Microbiol Biotechnol. 2022;106(19):6365-81.
  • Francolini I, Norris P, Piozzi A, Donelli G, Stoodley P. Usnic acid, a natural antimicrobial agent able to inhibit bacterial biofilm formation on polymer surfaces. Antimicrob Agents Chemother. 2004;48(11):4360-5.
  • Nithyanand P, Beema Shafreen RM, Muthamil S, Karutha Pandian S. Usnic acid, a lichen secondary metabolite inhibits Group A Streptococcus biofilms. Antonie Van Leeuwenhoek. 2015;107:263-72.
  • Grumezescu AM, Cotar AI, Andronescu E, Ficai A, Ghitulica CD, Grumezescu V, et al. In vitro activity of the new water-dispersible Fe₃O₄@usnic acid nanostructure against planktonic and sessile bacterial cells. J Nanopart Res. 2013;15:1766.
Toplam 34 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Konular Eczacılık ve İlaç Bilimleri (Diğer)
Bölüm Araştırma Makaleleri
Yazarlar

Ilker Kiliccioglu 0000-0003-3367-9665

Ayşegül Akkoyunlu 0000-0002-3663-7412

Gorkem Dulger 0000-0002-1506-1549

Başaran Dülger 0000-0002-3184-2652

Yayımlanma Tarihi 24 Eylül 2025
Gönderilme Tarihi 27 Mart 2025
Kabul Tarihi 1 Temmuz 2025
Yayımlandığı Sayı Yıl 2025 Cilt: 15 Sayı: 3

Kaynak Göster

Vancouver Kiliccioglu I, Akkoyunlu A, Dulger G, Dülger B. Usnea longissima Ach. Etanolik Özütünün HCT116 Kolon Kanseri Hücrelerinde Antikanser Etkisi ve Biyofilm İnhibisyon Kapasitesi. SABD. 2025;15(3):409-16.