Kukurbitasin D ve I'nın sorafenib ile kombinasyonunun hepatoselüler karsinoma hücre hattında RAS/RAF/MEK/ERK sinyal yolu aracılı apoptotik etkilerinin araştırılması
Küçük Resim Yok
Tarih
2022
Yazarlar
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Cilt Başlığı
Yayıncı
İnönü Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Amaç: Multi-tirozin kinaz inhibitörü olan sorafenib, HCC tedavisinde birinci basamak tedavi olarak kullanılan sistemik bir ilaçtır. Genel sağkalımı arttırmak için sorafenibin etki gücünü arttıracak ve ilaç direncini engelleyecek kombine ilaç tedavilerinin geliştirilmesi gerekmektedir. Kukurbitasinler, birçok kanser türünde aşırı aktive olan sinyal yollarını baskılayarak antikanser etki gösteren doğal bileşiklerdir. Bu çalışmada sorafenibin etkinliğini arttırmak ve tedavi dozunu azaltmak amacıyla CuD ve CuI'nın sorafenible kombinasyonunun HepG2 hücre hattı üzerindeki antikarsinojenik etkileri araştırıldı. Materyal ve Metot: CuD ve CuI HepG2 hücrelerine tek başına veya sorafenible kombine uygulanarak hücre inhibisyonu, göçü, apoptozu, hücre döngüsü dağılımı, mitokondriyal membran potansiyeli (??m) incelendi. Ayrıca sorafenibin etki ettiği bilinen sinyal yollarına kukurbitasinlerin nasıl etki ettiğini ortaya koymak amacıyla; VEGF, EGFR, MMP-2, kaspaz kaskatı, PI3K/AKT/mTOR, Raf/MEK/ERK sinyal yollarıyla ilgili proteinler ve genler western blot ve qRT-PCR ile değerlendirildi. Bulgular: CuD ve CuI'nın düşük konsantrasyonlarda antiproliferatif etkiye sahip olduğu, sorafenib ile kombine edildiğinde sinerjistik etki gösterdiği ortaya konmuştur. Kombine uygulamalar, kaspaz?9, Bax aktivitesini artırıp Bcl?xL aktivasyonunu inhibe ederek, hücre döngüsünü G2/M fazında bloke etti ve ??m kaybına neden olarak apoptoz indüksiyonunu sağladı. Ayrıca kombinasyonlar, hücre göçünü azaltarak, MMP-2 ve VEGF'i baskıladı. Kombine kukurbitasin-sorafenib uygulamaları EGFR ve PI3K/AKT/mTOR, Raf/MEK/ERK sinyallerinde rol alan proteinlerin ve genlerin ekspresyonunu inhibe etti. Sonuç: Kukurbitasinlerin sorafenible sinerjetik etki göstermesi ve benzer sinyal yollarını hedeflemesi, kombinasyonlarının HCC'de anjiyojenik, metastatik ve proliferatif aktiviteyi baskılayarak sorafenibin etkinliğini arttırabileceğini ortaya koymuştur.
Aim: Sorafenib, a multi-tyrosine kinase inhibitor, is a systemic drug used as first-line therapy in the treatment of HCC. In order to increase overall survival, it is necessary to develop combined drug therapies that will increase the potency of sorafenib and prevent drug resistance. Cucurbitacins are natural compounds that show anticancer effects by suppressing over-activated signaling pathways in many cancer types. In this study, the anticarcinogenic effects of sorafenib-CuD and CuI combination on HepG2 cell line were investigated in order to increase the efficacy of sorafenib and decrease the treatment dose. Material and Method: Cell inhibition, migration, apoptosis, cell cycle distribution, mitochondrial membrane potential (??m) were investigated by applying CuD and CuI alone or in combination with sorafenib to HepG2 cells. In addition, in order to reveal how cucurbitacins affect the signal pathways known to affect sorafenib; proteins and genes involved in VEGF, EGFR, MMP-2, caspase cascade, PI3K/AKT/mTOR, Raf/MEK/ERK signaling pathways were assessed by western blot and qRT-PCR. Results: It has been shown that CuD and CuI have an antiproliferative effect at low concentrations and show a synergistic effect when combined with sorafenib. Combined administrations induced apoptosis by increasing caspase-9, Bax activity and inhibiting Bcl?xL activation, blocking the cell cycle in G2/M phase and causing loss of ??m. The combinations also suppressed MMP-2 and VEGF, reduced cell migration. The combined cucurbitacin-sorafenib applications inhibited the expression of proteins and genes involved in EGFR and PI3K/AKT/mTOR, Raf/MEK/ERK signaling pathways. Conclusion: Due to showing the synergetic effect of cucurbitacins with sorafenib and their targeting of similar signaling pathways reveal that their combination may increase the efficacy of sorafenib by suppressing angiogenic, metastatic and proliferative activity in HCC.
Aim: Sorafenib, a multi-tyrosine kinase inhibitor, is a systemic drug used as first-line therapy in the treatment of HCC. In order to increase overall survival, it is necessary to develop combined drug therapies that will increase the potency of sorafenib and prevent drug resistance. Cucurbitacins are natural compounds that show anticancer effects by suppressing over-activated signaling pathways in many cancer types. In this study, the anticarcinogenic effects of sorafenib-CuD and CuI combination on HepG2 cell line were investigated in order to increase the efficacy of sorafenib and decrease the treatment dose. Material and Method: Cell inhibition, migration, apoptosis, cell cycle distribution, mitochondrial membrane potential (??m) were investigated by applying CuD and CuI alone or in combination with sorafenib to HepG2 cells. In addition, in order to reveal how cucurbitacins affect the signal pathways known to affect sorafenib; proteins and genes involved in VEGF, EGFR, MMP-2, caspase cascade, PI3K/AKT/mTOR, Raf/MEK/ERK signaling pathways were assessed by western blot and qRT-PCR. Results: It has been shown that CuD and CuI have an antiproliferative effect at low concentrations and show a synergistic effect when combined with sorafenib. Combined administrations induced apoptosis by increasing caspase-9, Bax activity and inhibiting Bcl?xL activation, blocking the cell cycle in G2/M phase and causing loss of ??m. The combinations also suppressed MMP-2 and VEGF, reduced cell migration. The combined cucurbitacin-sorafenib applications inhibited the expression of proteins and genes involved in EGFR and PI3K/AKT/mTOR, Raf/MEK/ERK signaling pathways. Conclusion: Due to showing the synergetic effect of cucurbitacins with sorafenib and their targeting of similar signaling pathways reveal that their combination may increase the efficacy of sorafenib by suppressing angiogenic, metastatic and proliferative activity in HCC.
Açıklama
Anahtar Kelimeler
Biyokimya, Biochemistry
