PANKREAS KANSERİ HÜCRELERİNDE 3-BROMOPÜRUVİK ASİDİN METABOLİK DEĞİŞİKLİKLER ÜZERİNDEKİ ANTİKANSER ETKİLERİNİN İN VİTRO OLARAK ARAŞTIRILMASI

Abstract

Amaç: Pankreatik duktal adenokarsinom (PDAC), yüksek mortalite oranı ve kemoterapötiklere karşı direnç ile karakterize edilen en agresif kanser türlerinden biridir. TP53, SMAD4 ve CDKN2A genlerindeki sık mutasyonlar, metabolik yeniden programlamayı tetikleyerek tümör progresyonuna ve tedaviye karşı direnç gelişimine katkıda bulunur. Glikolizin kilit enzimlerinden biri olan Heksokinaz 2 (HK2), PDAC'da sıklıkla aşırı eksprese edilir ve umut verici bir terapötik hedef olarak öne çıkar. Bu çalışma, TP53, SMAD4 ve CDKN2A mutasyonlarını taşıyan BXPC-3 PDAC hücrelerinde 3-bromopürivik asidin (3-BrPA) metabolik yollar, oksidatif stres ve ferroptozis üzerindeki antikanser etkilerini araştırmayı amaçlamıştır. Gereç ve Yöntemler: BXPC-3 hücreleri farklı konsantrasyonlarda 3-BrPA ile muamele edilmiştir. Hücre canlılığı, IC₅₀ değerinin belirlenmesi amacıyla MTT testi ile değerlendirilmiştir. Morfolojik değişiklikler ışık mikroskobu ile incelenmiştir. HK2 ekspresyonu ile birlikte GST, G6PD, 6-PGD ve GR enzimlerinin aktiviteleri ölçülmüş; glutatyon redoks durumu ve hücre içi demir seviyeleri değerlendirilmiştir. Ayrıca, yağ asidi oksidasyonu yoluyla metabolik kompansasyonu değerlendirmek üzere CPT1C aktivitesi analiz edilmiştir. Bulgular: 3-BrPA uygulaması, doz bağımlı şekilde hücre canlılığını anlamlı düzeyde azaltmış ve HK2 ekspresyonunu baskılamıştır. Glikoliz ve pentoz fosfat yolunun enzimatik aktiviteleri inhibe edilmiş; antioksidan kapasite azalmış ve oksidatif stres belirteçleri artmıştır. Bu değişiklikler, hücre içi demir düzeylerinde artış ve GSH/GSSG oranında azalma ile birlikte gözlenmiş; ferroptozis indüksiyonunu işaret etmiştir. Ayrıca, CPT1C aktivitesinde artış saptanmıştır, bu da yağ asidi oksidasyonuna yönelik bir metabolik kaymaya işaret etmektedir. Sonuç: Elde edilen bulgular, 3-BrPA’nın HK2 inhibisyonu yoluyla glukoz metabolizmasını bozduğunu, oksidatif stresi artırarak ferroptozisi tetiklediğini ortaya koymaktadır. Bu çalışma, 3-BrPA’nın kanser metabolizması ve ferroptozisi hedefleyen terapötik potansiyeline dikkat çekmektedir.

Keywords

• Pankreatik duktal adenokarsinom
• 3-bromopirüvik asit
• heksokinaz 2 inhibitörleri
• ferroptozis
• oksi datif stres
• pentoz fosfat yolu

INVESTIGATING THE ANTICANCER EFFECTS OF 3-BROMOPYRUVIC ACID ON METABOLIC ALTERATIONS IN PANCREATIC CANCER CELLS IN VITRO

Abstract

Objective: Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive cancers, characterised by a high mortality rate and resistance to chemotherapeutics. Frequent mutations in TP53, SMAD4, and CDKN2A drive metabolic reprogramming and contribute to tumour progression and treatment resistance. Hexokinase 2 (HK2), a key glycolytic enzyme, is commonly overexpressed in PDAC and represents a promising therapeutic target. This study aimed to investigate the anticancer effects of 3-bromopyruvic acid (3-BrPA) on metabolic pathways, oxidative stress, and ferroptosis in BXPC-3 PDAC cells harbouring TP53, SMAD4, and CDKN2A mutations. Material and Methods: BXPC-3 cells were administered with various concentrations of 3-BrPA, and cell viability was assessed using the MTT assay to determine the IC₅₀. The morphological changes were evaluated via light microscopy. HK2 expression and the enzymatic activities of GST, G6PD, 6-PGD, and GR were measured, along with the glutathione redox status and intracellular iron levels. CPT1C activity was additionally assessed to evaluate the metabolic alterations associated with the fatty acid oxidation pathways. Results: 3-BrPA administration significantly decreased cell viability in a dose-dependent manner and downregulated HK2 expression. It inhibited glycolysis and the pentose phosphate pathway enzymes, reduced the antioxidant capacity, and elevated oxidative stress markers. These changes were associated with increased intracellular iron levels and decreased GSH/GSSG ratios, indicating the induction of ferroptosis. Furthermore, CPT1C activity was upregulated, indicating increased fatty acid oxidation. Conclusion: Our findings demonstrate that 3-BrPA disrupts glucose metabolism, induces oxidative stress, and triggers ferroptosis in PDAC cells through HK2 inhibition. This study highlights 3-BrPA’s therapeutic potential in targeting cancer metabolism and ferroptosis.

Keywords

• Pancreatic ductal adenocarcinoma
• 3-bromopyruvic acid
• hexokinase 2 inhibitors
• ferroptosis
• oxidative stress
• pentose phosphate pathway

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