Activation of the Parthanatos Signaling Pathway by the Synergistic Combination of Cisplatin and Resveratrol in Tongue Cancer Cells

Öz

Tongue cancer is an aggressive oral malignancy with limited treatment success due to high invasion rates and chemotherapeutic resistance. Despite conventional therapies like surgery and radiotherapy, resistance to cornerstone drugs such as cisplatin remains a major obstacle, negatively impacting patient survival. Combining anti-cancer agents with distinct mechanisms is a critical strategy to overcome resistance and prevent tumor recurrence. Cisplatin induces DNA damage and apoptosis but is limited by systemic toxicity and acquired resistance. Resveratrol, a natural polyphenol with potent anticancer and antioxidant properties, has shown efficacy against various cancers, including multidrug-resistant cells. This study comprehensively investigated the synergistic cytotoxic effects of the cisplatin and resveratrol combination on the OSC-19 human tongue cancer cell line and its molecular relationship with the parthanatos cell death pathway. Cell viability was evaluated using the MTT assay, revealing that the combination therapy significantly inhibited proliferation compared to single-agent treatments, generating a potent synergistic response. To elucidate the molecular mechanism, mRNA expression levels of PARP-1 and AIF, key components of the parthanatos pathway, were analyzed via real-time PCR (RT-PCR). Findings demonstrated that the combination markedly triggered PARP-1 activation and upregulated AIF expression, thereby activating the parthanatos pathway. Distinct from classical apoptosis, parthanatos is a caspase-independent cell death form characterized by PARP-1 hyperactivation and the translocation of AIF from the mitochondria to the nucleus. This study provides scientific evidence that the cisplatin and resveratrol combination represents a novel and effective therapeutic approach to overcoming chemotherapy resistance in tongue cancer cells by inducing parthanatos.

Anahtar Kelimeler

• Human Tongue Cancer
• Cisplatin
• Resveratrol
• Synergistic Effect
• PARP-1

İnsan Dil Kanseri Hücrelerinde Sisplatin ve Resveratrol Sinerjistik Kombinasyonu ile Parthanatos Sinyal Yolağının Aktivasyonu

Öz

Dil kanseri, dünya genelinde yaygın ve tedavi başarısı sınırlı bir oral kanser türüdür. Cerrahi, radyoterapi ve kemoterapi gibi mevcut tedavilere rağmen, yüksek invazyon ve metastaz oranları ile kemoterapötik ilaçlara karşı gelişen direnç, hastaların yaşam kalitesini ve sağkalımını olumsuz etkilemektedir. Özellikle sisplatin gibi temel kemoterapötik ajanlara karşı gelişen direnç, tedavi önündeki en büyük engellerden biridir. Bu direncin üstesinden gelmek için farklı etki mekanizmalarına sahip anti-kanser ajanlarının kombinasyon halinde kullanılması kritik bir stratejidir; zira tek ilaç tedavileri sıklıkla dirençli mutasyonlara ve tümör nüksetmesine yol açabilmektedir. Sisplatin, DNA hasarı ve apoptoz indüksiyonu yoluyla etki gösteren güçlü bir kemoterapötiktir, ancak yan etkileri ve direnç gelişimi kullanımını kısıtlar. Resveratrol ise üzüm gibi bitkilerde bulunan, antikanser ve antioksidan özelliklere sahip doğal bir mikrobesindir. Çeşitli kanser türlerinde anti-proliferatif ve pro-apoptotik aktivite gösterdiği, ayrıca çoklu ilaç direncine sahip tümör hücrelerine karşı da etkili olduğu kanıtlanmıştır. Bu çalışmada, sisplatin ve resveratrol kombinasyonunun OSC-19 insan dil kanseri hücre hattı üzerindeki sinerjistik sitotoksik etkileri ve bu etkilerin moleküler düzeyde parthanatos hücre ölümü yolağı ile olan ilişkisi kapsamlı bir şekilde araştırılmıştır. Hücre canlılığı üzerindeki etkiler MTT testi ile değerlendirilmiş; elde edilen veriler kombinasyon tedavisinin, tekli ilaç uygulamalarına kıyasla hücre proliferasyonunu anlamlı düzeyde inhibe ettiğini ve güçlü bir sinerjistik sitotoksik yanıt oluşturduğunu ortaya koymuştur. Moleküler mekanizmayı aydınlatmak amacıyla, parthanatos yolağının kilit bileşenleri olan PARP-1 ve AIF genlerinin mRNA ekspresyon düzeyleri gerçek zamanlı polimeraz zincir reaksiyonu (RT-PCR) yöntemi ile analiz edilmiştir. Bulgular, kombinasyon tedavisinin PARP-1 aktivasyonunu belirgin şekilde tetiklediğini ve buna bağlı olarak AIF ekspresyonunu yukarı regüle ederek parthanatos yolağını aktive ettiğini göstermiştir. Parthanatos, klasik apoptozdan farklı olarak PARP-1 hiperaktivasyonu ve AIF'in mitokondriden çekirdeğe translokasyonu ile karakterize edilen, kaspaz bağımsız bir hücre ölümü formudur. Bu çalışma, sisplatin ve resveratrol kombinasyonunun dil kanseri hücrelerinde parthanatosu indükleyerek kemoterapi direncini kırmada ve tedavi etkinliğini artırmada yeni ve etkili bir terapötik yaklaşım olabileceğini bilimsel verilerle ortaya koymaktadır.

Anahtar Kelimeler

• İnsan Dil Kanseri
• Sisplatin
• Resveratrol
• Sinerjistik Etki
• PARP-1

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