Comparison of Epigenetic Changes of Endoplasmic Reticulum Stress and Autophagic Pathways in Various Cancer Cell Lines

Year 2025, Volume: 13 Issue: 1, 169 - 180

Nazmi Özgen , Şengül Yüksel

https://doi.org/10.33715/inonusaglik.1544914

Abstract

Disruption of the balance between processed protein load and protein folding capacity in the endoplasmic reticulum (ER), where post-translational modifications occur, results in the occurrence of adaptive ER stress, leading to the expectation of active autophagy. The relationship between ER stress and autophagic dysfunctions with cancer remains uncertain. The aim of this study is to investigate methylation changes in key genes in various cancer cells within these pathways. In this study, methylation changes in some major genes involved in ER stress and autophagic pathways were investigated in various cancer cells (HepG2, MCF-7) and healthy cells (MCF-10A) using the RT-PCR technique. Our study found that in MCF7 cells, the DNA methylation levels of ER stress genes MAP3K5, ATF6, and MAOA significantly decreased, while they increased in the XBP1 and EIF2AK3 genes. In HEPG2 cells, hypomethylation was detected in MAP3K5, EIF2AK3, and MAOA genes. Only GBA gene showed significant hypomethylation in both cells among autophagy pathway genes. With these results, it can be speculated that the epigenetic activities of most protein genes in ER stress and autophagic pathways are altered in cancer cells, thus accelerating the transformation process of cells.

Keywords

Autophagy, Cancer, DNA Methylation, ER stress, HepG2, MCF-7, MCF-10A

KANSER HÜCRELERİNDE ER STRESİ VE OTOFAJİK YOLAKLARDAKİ GENLERİN EPİGENETİK DEĞİŞİKLİKLERİNİN ARAŞTIRILMASI

Abstract

Post translasyonel modifikasyonların gerçekleştiği endoplazmik retikulumda (ER) işlenmiş protein yükü ile protein katlanma kapasitesi arasındaki dengenin bozulması sonucu, adaptif ER stresi oluşur ve otofajinin aktif olması beklenir. ER stresi ve otofajik bozuklukların kanser ile ilişkisi belirsizliğini korumaktadır. Bu çalışmada çeşitli kanser hücrelerinde bu yolaklardaki anahtar genlerdeki metilasyon değişikliklerinin araştırılması amaçlanmıştır. Çalışmada çeşitli kanser hücrelerinde (HepG2, MCF-7) ve sağlıklı hücrelerdeki (MCF-10A) ilgili genlerin metilasyon düzeyleri RT-PZR tekniği ile araştırıldı. Çalışmamızda MCF7 hücrelerinde ER stresi genlerinden MAP3K5, ATF6 ve MAOA’nin DNA metilasyon düzeylerinin anlamlı şekilde azaldığı XBP1 ve EIF2AK3 genlerinde ise arttığı belirlendi. HEPG2 hücrelerinde ise MAP3K5, EIF2AK3 ve MAOA genlerinde hipometilasyon saptandı. Otofaji yolağı genlerinden ise sadece GBA geninde her iki hücrede anlamlı hipometilasyon olduğu belirlendi. Bu sonuçlarla, kanser hücrelerinde ER stresi ve otofajik yolaklardaki çoğu protein genlerinin epigenetik olarak aktivitelerinin değiştirildiği böylece hücrelerin transformasyon sürecini hızlandırıldığı söyleyebiliriz.

Keywords

DNA Metilasyonu, ER stresi, HepG2, Kanser, Otofaji, MCF-7, MCF-10A

Supporting Institution

BAP

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