Doksorubisin ile Tedavi Edilen MCF-7 Meme Kanseri Hücrelerinde Melatonin ve Fukoidanın Oksidatif Stres ve Apoptoz Üzerindeki Farklılaştırıcı Etkileri

Öz

Amaç: Doksorubisin (DOX), östrojen reseptörü (ER)-pozitif meme kanserinin tedavisinde kullanılan temel kemoterapötik ajanlardan biridir. Sitotoksik etkileri büyük ölçüde mitokondriyal reaktif oksijen türleri (ROS) üretimi ve mitokondriye bağımlı apoptozun aktivasyonu aracılığıyla gerçekleşir. Bununla birlikte, seçici redoks modülatörlerinin DOX ile tetiklenen mitokondriyal stres yanıtlarını nasıl etkilediği ve bu etkilerin ER-bağımlı sinyal yolaklarıyla kesişip kesişmediği tam olarak açıklığa kavuşturulmamıştır. Bu çalışmada, melatonin ve fukoidanın DOX kaynaklı oksidatif stres ve apoptoz üzerindeki etkilerinin ER-pozitif MCF-7 meme kanseri hücrelerinde karşılaştırılması amaçlanmıştır. Materyal ve Metot: MCF-7 hücreleri 24 saat boyunca DOX (1 µM), melatonin (1 mM), fukoidan (100 µg/mL) veya bunların kombinasyonlarına maruz bırakıldı. Hücre canlılığı MTT yöntemi ile değerlendirildi. ROS düzeyleri DCFH-DA floresansı ile ölçülerek kontrol grubuna göre normalize edildi. Bax, Bcl-2, aktif (cleaved) kaspaz-3 ve ERα protein ekspresyonları ELISA yöntemi ile kantitatif olarak analiz edildi. Bulgular: DOX uygulaması hücre canlılığını anlamlı düzeyde azaltırken ve ROS üretimini artırırken (p<0.001), Bax ve aktif kaspaz-3 düzeylerini artırmış, Bcl-2 ve ERα ekspresyonunu azaltmış ve Bax/Bcl-2 dengesini pro-apoptotik yönde değiştirmiştir (p<0.001). Hem melatonin hem de fukoidan DOX kaynaklı ROS üretimini belirgin şekilde azaltmıştır (p<0.001); ancak her iki ajan da hücre canlılığını tam olarak geri kazandıramamış ve apoptotik belirteçleri normalize edememiştir. Fukoidan, melatonine kıyasla daha güçlü bir ROS baskılayıcı etki göstermiştir (p=0.021). Buna karşılık melatonin, ERα protein düzeylerini kısmen geri kazandırmış (p=0.008), fukoidan ise ERα ekspresyonu üzerinde anlamlı bir etki göstermemiştir. Sonuç: Melatonin ve fukoidan, ER-pozitif meme kanseri hücrelerinde DOX ile indüklenen oksidatif stres ve apoptotik yolakları farklı şekillerde modüle etmektedir. Bu bulgular, kemoterapiye bağlı hücresel stres yanıtlarının düzenlenmesinde mekanizma-spesifik ve birbirinden ayrışan rollerini ortaya koymaktadır.

Anahtar Kelimeler

• Doksorubisin
• Melatonin
• Fukoidan
• Oksidatif Stres
• Östrojen Reseptör Alfa

Destekleyen Kurum

Yok.

Etik Beyan

Bu çalışma kapsamında tüm deneyler yalnızca ticari olarak temin edilen insan hücre hattı üzerinde gerçekleştirildiğinden ve insan katılımcıları veya hayvan deneyleri içermediğinden etik kurul onayı gerekmemektedir.

Teşekkür

Yok.

Differential Modulation of Oxidative Stress and Apoptosis by Melatonin and Fucoidan in Doxorubicin-Treated MCF-7 Breast Cancer Cells

Öz

Objective: Doxorubicin (DOX) is a primary chemotherapeutic agent for estrogen receptor (ER)-positive breast cancer. Its cytotoxic effects are largely mediated by mitochondrial reactive oxygen species (ROS) production and the activation of mitochondria-dependent apoptosis. However, it remains unclear how selective redox modulators influence DOX-triggered mitochondrial stress responses and whether these effects intersect with ER-dependent signaling. This study aimed to compare the impacts of melatonin and fucoidan on DOX-induced oxidative stress and apoptosis in ER-positive MCF-7 breast cancer cells. Materials and Methods: MCF-7 cells were exposed for 24 hours to DOX (1 µM), melatonin (1 mM), fucoidan (100 µg/mL), or their combinations. Cell viability was assessed using the MTT assay. ROS levels were measured via DCFH-DA fluorescence and normalized to controls. Protein expression of Bax, Bcl-2, cleaved caspase-3, and ERα was quantified by ELISA. Results: DOX significantly decreased cell viability and elevated ROS production (p<0.001), while increasing Bax and cleaved caspase-3 levels, reducing Bcl-2 and ERα expression, and shifting the Bax/Bcl-2 balance toward a pro-apoptotic state (p<0.001). Both melatonin and fucoidan markedly attenuated DOX-induced ROS generation (p<0.001), although neither fully restored viability or normalized apoptotic markers. Fucoidan exhibited a stronger ROS-suppressive effect than melatonin (p=0.021). In contrast, melatonin partially restored ERα protein levels (p=0.008), whereas fucoidan showed no significant effect on ERα expression. Conclusions: Melatonin and fucoidan differentially modulate DOX-induced oxidative stress and apoptotic pathways in ER-positive breast cancer cells, highlighting distinct, mechanism-specific roles in regulating chemotherapy-associated cellular stress responses.

Anahtar Kelimeler

• Doxorubicin
• Melatonin
• Fucoidan
• Oxidative Stress
• Estrogen Receptor Alpha

Destekleyen Kurum

None.

Etik Beyan

Ethics committee approval was not required for this study, as all experiments were performed solely on commercially available human cell line and did not involve human subjects or animal experimentation.

Teşekkür

None.

Kaynakça

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