Exploring the Antiproliferative Mechanisms of Urtica dioica L. extract in Human Promyelocytic Leukemia Cell Line

Öz

Background: Nature is a source of therapeutic compounds which have fewer side effects compared to synthetic drugs. Stinging nettle extract, widely-used in Anatolia, has a strong antiproliferative effect on many types of cancer. However, the underlying molecular mechanisms of this effect is still not known. Materials and Methods: In this study, the interaction of Urtica dioica L. extract at different concentrations with apoptosis and autophagy pathways in the human promyelocytic cell line (HL-60) was studied to elucidate how it triggers the antiproliferative effects. In this context, firstly, the plant leaves were extracted in water with the Soxhlet extraction method. HL-60 cells were incubated with the extract at different concentrations for 24 hours, and the activated antitumoral effect pathway was investigated with advanced following molecular tests: MTT staining, Nitric oxide (NO) level, Annexin-V in flow cytometry, cell cycle, mitochondrial membrane potential measurement (MMP) and qPCR for evaluation of apoptosis and autophagy mediator genes. Results: It was determined that cell proliferation was suppressed at a concentration of 100 µg/mL and cells were kept in G0/G1 phase, MMP was impaired in cells and the rate of apoptotic cells increased. These apoptotic markers were confirmed by statistically increased expression of apoptotic and autophagy genes and NO level. Conclusions: Taken together, it is predicted that Urtica dioica L. water extract initiates apoptosis in HL-60 cells and could be promising compound candidate for cancer treatment.

Anahtar Kelimeler

• HL-60
• Urtica dioica
• Cancer
• Apoptosis
• APL

İnsan Promiyelositik Lösemi Hücre Hattında Isırgan Ekstraktının Antiproliferatif Mekanizmalarının Araştırılması

Öz

Amaç: Doğa, sentetik ilaçlara kıyasla daha az yan etkiye sahip terapötik bileşiklerin kaynağıdır. Anadolu'da yaygın olarak kullanılan ısırgan otu özütü, birçok kanser türü üzerinde güçlü bir antiproliferatif etkiye sahiptir. Bununla birlikte, bu etkinin altında yatan moleküler mekanizmalar hala bilinmemektedir. Materyal ve Metod: Bu çalışmada, insan promiyelositik hücre hattında (HL-60) farklı konsantrasyonlarda hazırlanan ısırgan ekstraktının apoptozis ve otofaji yolakları ile etkileşimi ve antiproliferatif etkileri incelenmiştir. Bu kapsamda öncelikle bitki yaprakları Soxhlet ekstraksiyon yöntemi ile suda ekstrakte edilmiştir. HL-60 hücreleri, 24 saat boyunca farklı konsantrasyonlarda ekstrakt ile inkübe edildi ve aktif anti-tümöral etki yolu, ileri moleküler testler ile araştırıldı: MTT boyama, Nitrik oksit (NO) seviyesi, flow sitometride Annexin-V, hücre döngüsü, mitokondriyal zar potansiyeli ölçümü (MMP) ile apoptozis ve otofaji yolak genlerinin değerlendirilmesi için qPCR. Bulgular: 100 µg/mL konsantrasyonda hücre proliferasyonunun baskılandığı ve hücrelerin G0/G1 fazında tutulduğu, hücrelerde MMP'nin bozulduğu ve apoptotik hücre yüzdesinin arttığı belirlendi. Bu apoptotik belirteçler, apoptotik ve otofaji genlerinin istatistiksel olarak artan ekspresyonu ve NO seviyesi ile doğrulandı. Sonuç: Tüm sonuçlar birlikte ele alındığında, ısırgan su ekstraktının HL-60 hücrelerinde apoptozisi başlattığı ve kanser tedavisi için umut verici bitki adayı olabileceği tahmin edilmektedir.

Anahtar Kelimeler

• HL-60
• Isırgan
• Kanser
• Apoptozis
• APL

Kaynakça

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