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

Abstract

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.

Keywords

• Usnea longissima Ach
• Colon cancer
• Caspase
• Anticancer
• Antibiofilm

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

Abstract

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.

Keywords

• Usnea longissima Ach
• Kolon kanseri
• Kaspaz
• Antikanser
• Antibiyofilm

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