Publication: Fisetin ile kombine edilmiş kemoterapi ilaçlarının kolorektal kanser hücrelerine etkisi üzerine çalışmalar
Abstract
Amaç: Kolorektal kanser hücrelerinde artan dozlarda kapesitabinle geliştirilen dirençli HT29 hücre soylarında yan etkileri yüksek olan kemoterapötik ajanının flavon fisetin ile kombine kullanılmasının hem kemoterapötik ilaçların toksik etkilerinin azaltması hem de apoptoz protein ekspresyonları ile spesifik miRNA ekspresyonlarının kolorektal kanser tedavisi üzerindeki etkisinin belirlenmesi amaçlanmıştır. Gereç ve Yöntem: Biyoinformatik analizler yapılarak kolorektal kanserlerle ilişkili miRNA’ lar belirlendikten sonra HT29 hücrelerinde kapesitabin ile artan dozlarda ilaç direnci geliştirilmiştir. Kapesitabin direnci geliştirilmiş hücreler fisetin ve/ veya kapesitabin ile inkübe edildikten sonra hücre çoğalması, hücre göçünün belirlenmesi, seçilen miRNA’ların ekspresyon değişimlerinin GZ-PZR çalışmaları ve apoptoz protein ekspresyon düzeyindeki değişimleri belirlemek üzere western emdirimi çalışmaları yapılmıştır. Bulgular: Kapesitabine dirençli HT29 hücre alt kümesi geliştirilmiştir. HT29 hücre soylarında geliştirilen kapesitabin direnci tüm deney sonuçları ile gösterilmiştir. Yabanıl HT29 ve kapesitabine dirençli HT29 hücrelerinde fisetinin tek başına ve/ veya kapesitabinle kombinasyonlarının hücre çoğalması, yara iyileşmesi, koloni oluşumu, miRNA ekspresyonları ve apoptoz protein ekspresyonlarında değişimlere neden olmuştur. Sonuç: Fisetinin, hem yabanıl-tip hem de kapesitabin direnci geliştirilmiş hücrelerde tek başına ve kapesitabin ile kombinasyonunun hücre çoğalması, üzerinde inhibe edici etkileri gösterilmiştir. Ayrıca yabanıl ve dirençli hücrelerde kapesitabin, fisetin ve kombiasyonları, miR 21 ekspresyonunda azalma ilaç duyarlılığının artmasına katkı sağlayabilir. miRNA’ lardaki değişimler tümör baskılanmasına ve direncin gerilemesine de katkıda bulunabilir.
Objective: It was aimed to use the chemotherapeutic agent, which has high side effects, in combination with the flavone fisetin, in resistant HT29 cell lines developed with increasing doses of capecitabine in colorectal cancer cells, to both reduce the toxic effects of chemotherapeutic drugs and to determine the effect of apoptosis protein expressions and specific miRNA expressions on colorectal cancer treatment. Material and Methods: After miRNAs associated with colorectal cancers were determined by bioinformatic analysis, drug resistance was developed in HT29 cells with increasing doses of capecitabine. After capecitabine-resistant cells were incubated with fisetin and/ or capecitabine, western blot studies were performed to determine cell proliferation, cell migration, RT-PCR studies of expression changes of selected miRNAs, and changes in the expression level of apoptosis proteins. Results: A subset of capecitabine-resistant HT29 cells has been developed. Capecitabine resistance developed in HT29 cell lines was demonstrated by all experimental results. In wild-type HT29 and capecitabine-resistant HT29 cells, fisetin alone and/ or in combination with capecitabine caused changes in cell proliferation, wound healing, colony formation, miRNA expressions and apoptosis protein expressions. Conclusion: Fisetin, alone and in combination with capecitabine, has been shown to have inhibitory effects on cell proliferation in both wild-type and capecitabine-resistant cells. Additionally, in wild-type and resistant cells, capecitabine, fisetin and their combinations may contribute to decreased miR-21 expression and increased drug sensitivity. Changes in miRNAs may also contribute to tumor suppression and regression of resistance.
Objective: It was aimed to use the chemotherapeutic agent, which has high side effects, in combination with the flavone fisetin, in resistant HT29 cell lines developed with increasing doses of capecitabine in colorectal cancer cells, to both reduce the toxic effects of chemotherapeutic drugs and to determine the effect of apoptosis protein expressions and specific miRNA expressions on colorectal cancer treatment. Material and Methods: After miRNAs associated with colorectal cancers were determined by bioinformatic analysis, drug resistance was developed in HT29 cells with increasing doses of capecitabine. After capecitabine-resistant cells were incubated with fisetin and/ or capecitabine, western blot studies were performed to determine cell proliferation, cell migration, RT-PCR studies of expression changes of selected miRNAs, and changes in the expression level of apoptosis proteins. Results: A subset of capecitabine-resistant HT29 cells has been developed. Capecitabine resistance developed in HT29 cell lines was demonstrated by all experimental results. In wild-type HT29 and capecitabine-resistant HT29 cells, fisetin alone and/ or in combination with capecitabine caused changes in cell proliferation, wound healing, colony formation, miRNA expressions and apoptosis protein expressions. Conclusion: Fisetin, alone and in combination with capecitabine, has been shown to have inhibitory effects on cell proliferation in both wild-type and capecitabine-resistant cells. Additionally, in wild-type and resistant cells, capecitabine, fisetin and their combinations may contribute to decreased miR-21 expression and increased drug sensitivity. Changes in miRNAs may also contribute to tumor suppression and regression of resistance.