İZOİMPERATORİN ARACILIKLI ANTİKANSER AKTİVİTE: MİTOKONDRİYAL DİSFONKSİYONUN HEPG2 HÜCRELERİNDEKİ ROLÜ

Öz

Amaç: Çalışma kapsamındaki ilk amaç, glukoz veya galaktoz içeren besiyerlerinde İzoimperatorin’e (İZO) maruz kalmış HepG2 hücrelerindeki mitokondrinin rolünü Crabtree etkisi nedeni ile araştırmaktır. İkinci amaç, mitokondriyal disfonksiyonun ortaya çıkmasında rolü olabilecek olan İZO ve elektron transfer zinciri arasındaki (ETZ) etkileşimi öngörmektir. Gereç ve Yöntem: Glukoz veya galaktoz içeren besiyerlerinde İZO (12.5, 25, 50, 100 ve 250 µM) ile inkübe edilen HepG2 hücrelerin canlılığı ve membran hasarı MTT ve LDH sızma deneyleri ile gerçekleştirilmiştir. İZO‘nun ETZ kompleksleri üzerine olan afinitesi moleküler kenetleme çalışması ile analiz edilmiştir. Sonuç ve Tartışma: MTT deneyi sonuçlarına göre glukoz içeren besiyerinde 250 µM İZO sitotoksik etki gösterirken, galaktoz içeren besiyerinde 25 µM ve daha yüksek konsantrasyonlar hücre canlılığını azaltmıştır. Glukoz içeren besiyerinde gerçekleştirilen membran hasarı deneyi, 250 uM İZO’nun membran bütünlüğünü bozduğunu göstermiştir. Galaktoz içeren besiyerinde 100 ve 250 uM İSO membran hasarını artırmıştır. Moleküler kenetlenme çalışma sonuçlarına göre İZO’nun ETZ komplekleri üzerine olan afinitesi Kompleks IV > Kompleks I > Kompleks III > Kompleks II şeklindedir. İZO, elektronun sitokrom C’den oksijene aktarılmasını engelleyerek kompleks IV’ün inhibisyonunu yapmakta ve proton gradiyentinin azalmasına neden olmaktadır. Elde edilen sonuçlar, İZO’nun gerçekleştirdiği ETZ inhibisyonunun mitokondriyal disfonksiyona neden olabileceğini göstermektedir.

Anahtar Kelimeler

• Antikanser
• crabtree etkisi
• elektron transfer zinciri
• izoimperatorin
• mitokondriyal disfonksiyon

ISOIMPERATORIN-MEDIATED ANTICANCER ACTIVITY: ROLE OF MITOCHONDRIAL DYSFUNCTION IN HEPG2 CELLS

Öz

Objective: The first goal of the present study is to investigate the role of mitochondria due to the Crabtree effect in HepG2 cells exposed to ISO in either glucose- or galactose-conditioned media. The second aim is to predict the interactions between electron transport chain (ETC) complexes and ISO, which might be the possible reason for mitochondrial dysfunction. Material and Method: Cell viability and membrane damage for HepG2 cells exposed to ISO (12.5, 25, 50, 100, and 250 µM) were assessed by MTT and LDH leakage assays in either glucose- or galactose-conditioned media. The affinity of ISO to ETC complexes was also determined by a molecular docking study. Result and Discussion: MTT assay showed that 250 µM ISO leads to cytotoxic activity in glucose-conditioned media, while 25 µM and higher concentrations of ISO decrease cell viability in galactose-conditioned media. A membrane damage assay conducted in a glucose-conditioned media assay revealed that 250 µM ISO disrupts the cell membrane. 100 and 250 µM ISO increased membrane damage in galactose-conditioned media. According to docking simulations, binding affinities of ISO to ETC complexes are in descending order: Complex IV > Complex I > Complex III > Complex II. Inhibition of complex IV by ISO inhibits the transfer of electrons from cytochrome c to oxygen, and the proton gradient collapses. The present study proposed that ISO leads to mitochondrial dysfunction via inhibition of the ETC.

Anahtar Kelimeler

• Anticancer
• crabtree effect
• electron transport chain
• isoimperatorin
• mitochondrial dysfunction

Teşekkür

We gratefully thank Prof. Ayşe NALBANTSOY (Bioengineering Department, Ege University) for the laboratory facilities.

Kaynakça

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