Kolorektal Kanser Hücrelerinde Boraksın Gpx4/ACSL4 Sinyal Yolu Aracılığıyla Sitotoksik Etkileri

Abstract

Amaç: Kolorektal kanser (CRC), kansere bağlı ölümlerin yaklaşık %10'unu oluşturmasıyla, malignite yönünden üçüncü ve kansere bağlı ölümlerde ikinci sırada yer almaktadır. Ferroptoz, CRC dahil çok sayıda kanserin tedavisinde ilaç direncini önlemede terapötiklerin anti-kanser etkinliğini arttırabilecek potansiyel demire bağlı hücre ölüm yolağıdır. Bu çalışmada CRC hücrelerinde ferroptozu sinyal yolağı üzerinden boraksın anti-proliferatif etkilerini araştırmak amaçlandı. Gereç ve Yöntemler: Öncelikle, boraksın sitotoksik konsantrasyonları (0-64 mM aralığında) 3-(4,5-dimetiltiazol-2-il)-2,5-difenil tetrazolyum bromür (MTT) testi ile belirlendi. Daha sonra sitotoksik boraks konsantrasyonları ile 24 saat inkübe edilen HCT-116 hücrelerinde glutatyon peroksidaz 4 (GPx4), açil-KoA sentetaz uzun zincirli aile üyesi 4 (ACSL4), malondialdehit (MDA) ve 8-hidroksideoksiguanozin (8-OHdG) seviyeleri belirlendi. Bulgular: 1 ve 4 mM boraks konsantrasyonları hücre canlılığını etkilemezken, 8 mM ve üzerindeki boraks konsantrasyonları HTC-116 hücrelerinde canlılığı anlamlı şekilde düşürmüştür (p<0,05). MTT sonuçlarına göre, boraksın IC25 ve IC50 konsantrasyonları sırasıyla 12,5 mM ve 20,8 mM olarak belirlendi. HTC-116 hücrelerinde 24 saat boyunca 12.5, 16,2 ve 20,8 mM boraks maruziyeti GPx4 seviyelerinde konsantrasyon bağlım şekilde düşüş olurken, ACSL4 seviyelerinde artış olmuştur. Ayrıca, HTC-116 hücrelerine artan boraks uygulamasının, kontrole kıyasla MDA ve 8-OHdG seviyelerinde önemli bir artış ile lipit peroksidasyonuna ve DNA hasarına neden olduğu bulundu (p<0,05). Sonuç: Deneysel verilerimiz doğrultusunda, boraks insan CRC hücrelerinde potansiyel bir anti-kanser ajanı olarak ferroptozu indükleyerek anti-proliferatif etkiler göstermiştir.

Keywords

• Boraks
• DNA hasarı
• ferroptoz
• HTC-116
• lipit peroksidasyonu

Cytotoxic Effects of Borax via GPx4/ACSL4 Signaling Pathway in Colorectal Cancer Cells

Abstract

Aim: Colorectal cancer (CRC) ranks third in terms of malignancy and second in cancer-related deaths, accounting for approximately 10% of cancer-related deaths. Ferroptosis is a potential iron-induced cell death pathway that could increase the anti-cancer efficacy of therapeutics in preventing drug resistance in the treatment of many cancers, including CRC. In this study, we aimed to investigate the anti-proliferative effects of borax via ferroptosis signaling pathway in CRC cells. Material and Methods: Firstly, we determined the cytotoxic concentrations of borax (range 0 to 64 mM) with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Subsequently, glutathione peroxidase 4 (GPx4), acyl-CoA synthetase long chain family member 4 (ACSL4), malondialdehyde (MDA) and 8-hydroxydeoxyguanosine (8-OHdG) levels were determined in HCT-116 cells incubated with cytotoxic borax concentrations for 24 hours. Results: Borax at 1 and 4 mM concentrations did not affect cell viability, while borax at 8 mM and above concentrations significantly reduced viability in HTC-116 cells (p<0.05). According to MTT results, IC25 and IC50 concentrations of borax were determined as 12.5 mM and 20.8 mM, respectively. In HTC-116 cells, exposure to 12.5, 16.2, and 20.8 mM borax for 24 hours resulted in a concentration-dependent decrease in GPx4 levels and an increase in ACSL4 levels. We also found that increased borax administration to HTC-116 cells caused lipid peroxidation and DNA damage with a significant increase in MDA and 8-OHdG levels compared to control (p<0.05). Conclusion: In line with our experimental data, borax showed anti-proliferative effects by inducing ferroptosis as a potential anti-cancer agent in human CRC cells.

Keywords

• Borax
• DNA damage
• ferroptosis
• HTC-116
• lipid peroxidation

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