Ferula orientalis Ekstraktı, MDA-MB-231 Üçlü Negatif Meme Kanseri Hücrelerinde Redoks Modülasyonu Yoluyla Mitokondriyal Apoptozu İndükler

Öz

Amaç: Üçlü negatif meme kanseri (TNBC), östrojen reseptörü, progesteron reseptörü ve HER2 ekspresyonunun olmaması ile karakterize edilen, agresif seyirli bir meme kanseri alt tipidir. Hedefe yönelik tedavi seçeneklerinin sınırlı olması ve tümör hücrelerinin redoks adaptasyonları nedeniyle TNBC hücreleri artmış oksidatif kırılganlık göstermektedir; bu durum redoks modülasyonunu umut verici bir terapötik strateji haline getirmektedir. Fitokimyasal açıdan zengin bir tıbbi bitki olan Ferula orientalis’in potansiyel antikanser özelliklere sahip biyoaktif bileşikler içerdiği bildirilmiştir. Bununla birlikte, TNBC hücrelerinde redoks homeostazı ve mitokondriyal apoptoz üzerindeki etkileri büyük ölçüde aydınlatılmamıştır. Bu çalışma, Ferula orientalis ekstraktının MDA-MB-231 TNBC hücre hattında oksidatif stres parametreleri, antioksidan enzim aktiviteleri ve apoptoz ile ilişkili gen ekspresyonu üzerindeki etkilerini araştırmayı amaçlamıştır.

Gereç ve Yöntemler: MDA-MB-231 hücreleri 24 saat süreyle 10, 50 ve 100 µg/mL konsantrasyonlarında F. orientalis ekstraktı ile muamele edilmiştir. Hücre içi indirgenmiş glutatyon (GSH) ve malondialdehit (MDA) düzeyleri redoks durumunu değerlendirmek amacıyla spektrofotometrik yöntemlerle ölçülmüştür. Süperoksit dismutaz (SOD), katalaz (CAT) ve glutatyon peroksidaz (GPX) aktiviteleri enzimatik analizlerle belirlenmiştir. Apoptozla ilişkili gen ekspresyon düzeyleri (BAX ve BCL2), 2^−ΔΔCt yöntemi kullanılarak RT-qPCR ile analiz edilmiştir. İstatistiksel değerlendirme tek yönlü ANOVA ve Tukey post-hoc testi ile yapılmıştır.

Bulgular: F. orientalis uygulaması GSH düzeylerinde doza bağımlı azalma ve MDA düzeylerinde anlamlı artışa yol açarak oksidatif stresin arttığını göstermiştir. Antioksidan enzim aktiviteleri (SOD, CAT ve GPX) tüm tedavi gruplarında anlamlı şekilde baskılanmıştır. Ayrıca BAX ekspresyonu artarken BCL2 ekspresyonu azalmış ve BAX/BCL2 oranı belirgin şekilde yükselmiştir. Bu bulgular, ekstraktın redoks dengesizliği üzerinden mitokondriyal apoptotik sinyalizasyonu aktive ettiğini göstermektedir.

Sonuç: Ferula orientalis ekstraktı TNBC hücrelerinde redoks homeostazını bozmakta ve oksidatif stres aracılı mitokondriyal apoptozu indüklemektedir. Oksidatif yükün artırılması ve anti-apoptotik savunma mekanizmalarının baskılanmasını içeren bu çift yönlü etki, F. orientalis’in TNBC’de redoks-hedefli tamamlayıcı terapötik bir aday olarak değerlendirilmesini desteklemektedir. Bulguların in vivo modellerde doğrulanması ve aktif bileşenlerin tanımlanması gerekmektedir.

Anahtar Kelimeler

• Üçlü negatif meme kanseri
• Ferula orientalis
• Oksidatif stres
• Redoks homeostazı
• Mitokondriyal apoptoz

Ferula orientalis Extract Induces Mitochondrial Apoptosis Through Redox Modulation in MDA-MB-231 Triple-Negative Breast Cancer Cells

Öz

Objective: Triple-negative breast cancer (TNBC) is an aggressive molecular subtype of breast cancer characterized by the absence of estrogen receptor, progesterone receptor, and HER2 expression. Due to limited targeted therapeutic options and intrinsic redox adaptations, TNBC cells exhibit increased oxidative vulnerability, making redox modulation a promising therapeutic strategy. Ferula orientalis, a phytochemically rich medicinal plant, has been reported to possess bioactive compounds with potential anticancer properties. However, its effects on redox homeostasis and mitochondrial apoptosis in TNBC cells remain largely unexplored. This study aimed to investigate the impact of Ferula orientalis extract on oxidative stress parameters, antioxidant enzyme activities, and apoptosis-related gene expression in the MDA-MB-231 TNBC cell line.

Materials and Methods: MDA-MB-231 cells were treated with F. orientalis extract at concentrations of 10, 50, and 100 µg/mL for 24 hours. Intracellular reduced glutathione (GSH) and malondialdehyde (MDA) levels were measured spectrophotometrically to assess redox status. Superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) activities were determined using enzymatic assays. Apoptosis-related gene expression levels (Bax and Bcl-2) were quantified by RT-qPCR using the 2^−ΔΔCt method. Statistical analyses were performed using one-way ANOVA followed by Tukey’s post hoc test.

Results: Treatment with F. orientalis resulted in a dose-dependent depletion of GSH and a significant elevation of MDA levels, indicating enhanced oxidative stress. Antioxidant enzyme activities (SOD, CAT, and GPX) were significantly suppressed across treatment groups. Furthermore, Bax expression was upregulated, whereas Bcl-2 expression was downregulated, leading to a marked increase in the Bax / Bcl-2 ratio. These findings collectively demonstrate activation of mitochondrial apoptotic signaling in response to extract-induced redox imbalance.

Conclusion: Ferula orientalis extract disrupts redox homeostasis and promotes oxidative stress–mediated mitochondrial apoptosis in TNBC cells. The dual mechanism involving enhancement of oxidative burden and suppression of anti-apoptotic defenses highlights the potential of F. orientalis as a redox-targeted complementary therapeutic candidate in TNBC. Further studies are required to identify active constituents and validate these findings in in vivo models.

Anahtar Kelimeler

• Triple-negative breast cancer
• Ferula orientalis
• Oxidative stress
• Redox homeostasis
• Mitochondrial apoptosis

Kaynakça

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