Turkiye Klinikleri Journal of Medical Sciences

.: ORIGINAL RESEARCH
Trabektedinin Meme Kanseri Kök Hücreleri Üzerindeki Sitotoksik ve Apoptotik Etkileri
Cytotoxic and Apoptotic Effects of Trabectedin on Breast Cancer Stem Cells
Harika ATMACA İLHANa, Burcu ÇAKARb, Merve İŞSEVENa, Burçak KARACAb
aBiyoloji Bölümü, Manisa Celal Bayar Üniversitesi Fen Edebiyat Fakültesi, Manisa, TÜRKİYE
bTıbbi Onkoloji BD, Ege Üniversitesi Tıp Fakültesi, İzmir, TÜRKİYE
Turkiye Klinikleri J Med Sci. 2019;39(4):422-9
doi: 10.5336/medsci.2019-64806
Article Language: TR
Full Text
ÖZET
 Amaç: Bu çalışmada, etkileri henüz meme kanseri kök hücrelerinde araştırılmamış olan trabektedinin (ET743, Yondelis®) sitotoksik ve apoptotik etkileri MCF-7 hücreleri ve bu hücrelerden izole edilen CD44+/CD24- meme kanseri kök hücreleri üzerinde karşılaştırmalı olarak araştırılmıştır. Gereç ve Yöntemler: CD44+/CD24- belirteçlerine sahip meme kanseri kök hücreleri MCF-7 hücre kültüründen akış sitometri metodu ile izole edildi. Trabektedinin MCF-7 ve CD44+/CD24- meme kanseri kök hücrelerinin canlılığı üzerindeki etkisini belirlemek üzere XTT testi kullanıldı. Trabektedinin, hücrelerin canlılığı üzerindeki etkisi konsantrasyona ve zamana bağlı olarak ölçüldü. Apoptotik etki DNA fragment miktarının ölçümüyle, mitokondriyal membran potansiyelinde meydana gelen değişimler ise tetrametilrodamin etil ester (TMRE) boyamasıyla belirlendi. Bulgular: Trabektedin, hücre canlılığını MCF-7 meme kanseri ve CD44+/CD24- meme kanseri kök hücre gruplarının her ikisinde de konsantrasyona ve zamana bağlı olarak inhibe etti. MCF-7 hücreleri 48. saatteki IC50 konsantrasyonları için 4,52±0,7 nM, CD44+/CD24- hücreleri için ise 1,23±1,2 nM olarak hesaplandı. IC50 konsantrasyonları değerlendirildiğinde, CD44+/CD24- meme kanseri kök hücrelerinin trabektedine MCF-7'den daha hassas olduğu tespit edildi. Kontrol grubundaki hücrelerin mitokondrileri floresans TMRE boyası ile boyanırken, trabektedinle muamele edilen her iki hücre popülasyonunun da boyanma miktarlarında azalma gözlendi. Trabektedinin CD44+/CD24- meme kanseri kök hücrelerinde apoptozu daha etkin bir şekilde indüklediği belirlendi. Sonuç: Elde edilen veriler, trabektedinin meme kanseri hücre grubunda olduğu gibi meme kanseri kök hücrelerinde de sitotoksik ve apoptotik etkisi olduğunu ortaya koymuştur. Bu sonuçlar, trabektedinin meme kanseri için potansiyel bir terapötik olabileceğini göstermektedir. Ancak trabektedinin meme kanseri kök hücreleri üzerindeki etkilerinin daha detaylı olarak çalışıldığı araştırmalara ihtiyaç vardır.

Anahtar Kelimeler: Trabektedin; meme kanseri; MCF-7; kanser kök hücresi; sitotoksisite; apoptozis
ABSTRACT
 Objective: In this study, effect of trabectedin (ET743, Yondelis®) was investigated on the viability of MCF-7 cells and CD44+/CD24- breast cancer stem cells. Material and Methods: CD44+/CD24- breast cancer stem cells were isolated from MCF-7 cells by using flow cytometry. XTT test was used to investigate the effect of trabectedin on the viability of MCF-7 and CD44+/ CD24- breast cancer stem cells. Apoptotic effect of trabectedin was investigated by measuring DNA fragmentation and changes in mitochondrial membrane potential were determined by tetramethylrhodamine ethyl ester (TMRE) staining. Results: Trabectedin inhibited the cell viability of both MCF-7 breast cancer and CD44+/CD24- breast cancer stem cells in a concentration and time dependent manner. IC50 concentrations at 48 h was calculated as 4.52±0.7 nM for MCF-7 cells and 1.23±1.2 nM for CD44+/CD24- cells. It was found that breast cancer stem cells were more sensitive to trabectedin than MCF-7 cancer cells. Cells in control group were stained with fluorescent TMRE stain but the staining was less in the both cell groups that were treated with the IC50 concentrations of trabectedin. It was shown that trabectedin induced more apoptotic cells in CD44+/CD24- breast cancer stem cells. Conclusion: According to the current data, it was shown that trabectedin induced cytotoxicity and apoptosis in breast cancer stem cells as well as MCF-7 cancer cells in a concentration and time dependent manner. These data indicates that trabectedin might be a potential therapeutic for breast cancer. However, more detailed studies investigating the effects of trabectedin on breast cancer stem cells are needed.

Keywords: Trabectedin; breast cancer; MCF-7; cancer stem cell; cytotoxicity; apoptosis
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