Fotoduyarlı biyomalzemelerle uygulanan fotodinamik terapi'nin epigenetik mekanizmalara etkisi
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Tarih
2022
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Yayıncı
Trakya Üniversitesi Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Fotodinamik terapi ile indüklenen, akut oksidatif strese maruz kalan hücrelerin, bu strese vereceği yanıtta genetik ve epigenetik değişikliklerin olası bağlantısına odaklandık. Fotoduyarlaştırıcı aracılı apoptozda yer alan mekanizmaları anlamak, terapötik etkinliğini artırmak bu yöntemin daha etkin kullanımını sağlayabilir. Bu çalışmanın amacı, HepG2 kanser hücrelerinde, ışık kaynağının farklı dozları kullanılarak, in vitro fotodinamik terapi uygulamasının mitokondriyal apoptoz genlerinin göreceli ekspresyonlarındaki değişim ve apoptoz yanıtından sorumlu moleküler, epigenetik mekanizmaları ve ajanların biyouyumlukları araştırıldı. Çalışmamızda üç farklı tipte fotoduyarlı ajandan elde ettiğimiz sonuçların fotodinamik terapi alanına biyomalzeme olarak katkı sağlayabileceğini düşünmekteyiz. Çalışmanın ilk kısmında, fotoduyarlı ajanların hücre canlılığına olan fototoksik etkisi test edildi. Ksenon ışık kaynağını kullanarak, ALA, Çinko ve Silisyum fotuduyarlaştırıcılar 600 mW/cm 2 'lik ışıma dozunda sırasıyla, 100 sn, 200 sn ve 300 sn süreler ile 0 saat, 24 saat ve 48 saat inkübasyon süreleri boyunca 10 farklı fotoduyarlı madde dozunda inkübe edilerek her bir zaman ve ışıma dozu için IC50 değerleri hesaplandı. Belirlenen bu IC50 değerlerinden 48 saat süre ile 100 sn ışıma dozu için hücrelerin apoptoz, genetik ve metilasyon ile ilgili gen analizleriyle ajanların biyomalzeme olup olmayacakları ilgili değerledirmelerde bulunuldu. Sitokrom c, Nrf2 ve Keap1'in epigenetik düzenlenmesinin, oksidatif stres veya ilaç metabolizması ile ilgili hastalıkların tedavisinde olası bir terapötik hedef olacağı düşünülmektedir. Bu malzemelerin, metilasyon inhibitör veya aktivatör adayları olup olamayacağı değerlendirildi. Bu malzemelerin ışıklı ve ışıksız olarak HepG2 hücrelerinde önemli miktarda apoptozu indüklediği gözlendi. Işıklı uygulamalarda apoptoz genlerinin ekspresyonunda, kontrol grubuna göre belli bir artış yaşandı. Sonuç olarak, tüm kullanılan fotoduyarlı biyomalzemelerin fototoksik, apoptotik ve epigenetik bir etkiye sahip olduklarını düşünmekteyiz.
We focused on the possible link between genetic and epigenetic changes in the response of cells exposed to acute oxidative stress induced by photodynamic therapy. Understanding the mechanisms involved in photosensitizer mediated apoptosis and increasing its therapeutic efficacy may enable more effective use of this method. The aim of this study was to investigate changes in relative expression of mitochondrial apoptosis genes and molecular, epigenetic mechanisms and bioavailability of drugs responsible for this apoptosis response in HepG2 cancer cells by using different doses of light source in vitro. We think that results obtained from three different types of photosensitive agents in our study may contribute to field of photodynamic therapy as a biomaterial. In the first part of the study, phototoxic effect of photosensitive agents on cell viability was tested. Using a xenon light source with ALA, Zinc and Silicon photosensitizers were incubated at an irradiation dose of 600 mW/cm 2 for 100 s, 200 s, and 300 s, respectively, for 0 hour, 24 hours, and 48 hours incubation times, with 10 different doses of photosensitive drugs. IC50 values were calculated for each time and each irradiation dose. From these determined IC50 values, it was evaluated whether the agents would be biomaterials with gene analyzes related to apoptosis, genetics and methylation for 48 hours and 100 sec irradiation dose. Epigenetic regulation of Cytochrome c, Nrf2 and Keap1 is thought to be a possible therapeutic target in the treatment of diseases related to oxidative stress or drug metabolism. It was evaluated whether these materials could be methylation inhibitor or activator candidates. It was observed that these materials induced a significant amount of apoptosis in HepG2 cells with and without light. There was a certain increase in the expression of apoptosis genes in light treatments compared to control group. In conclusion, we think that all used photosensitive biomaterials have a phototoxic, apoptotic, and epigenetic effect.
We focused on the possible link between genetic and epigenetic changes in the response of cells exposed to acute oxidative stress induced by photodynamic therapy. Understanding the mechanisms involved in photosensitizer mediated apoptosis and increasing its therapeutic efficacy may enable more effective use of this method. The aim of this study was to investigate changes in relative expression of mitochondrial apoptosis genes and molecular, epigenetic mechanisms and bioavailability of drugs responsible for this apoptosis response in HepG2 cancer cells by using different doses of light source in vitro. We think that results obtained from three different types of photosensitive agents in our study may contribute to field of photodynamic therapy as a biomaterial. In the first part of the study, phototoxic effect of photosensitive agents on cell viability was tested. Using a xenon light source with ALA, Zinc and Silicon photosensitizers were incubated at an irradiation dose of 600 mW/cm 2 for 100 s, 200 s, and 300 s, respectively, for 0 hour, 24 hours, and 48 hours incubation times, with 10 different doses of photosensitive drugs. IC50 values were calculated for each time and each irradiation dose. From these determined IC50 values, it was evaluated whether the agents would be biomaterials with gene analyzes related to apoptosis, genetics and methylation for 48 hours and 100 sec irradiation dose. Epigenetic regulation of Cytochrome c, Nrf2 and Keap1 is thought to be a possible therapeutic target in the treatment of diseases related to oxidative stress or drug metabolism. It was evaluated whether these materials could be methylation inhibitor or activator candidates. It was observed that these materials induced a significant amount of apoptosis in HepG2 cells with and without light. There was a certain increase in the expression of apoptosis genes in light treatments compared to control group. In conclusion, we think that all used photosensitive biomaterials have a phototoxic, apoptotic, and epigenetic effect.
Açıklama
Anahtar Kelimeler
Fotodinamik terapi, Apoptoz, Ftalosiyanin, Aminolevulinik Asit, Metilasyon, Photodynamic therapy, Apoptosis, Phthalocyanine, Aminolevulinic Acid, Methylation
