Akrilamidin insan kaynaklı hepatoma hücrelerinde protein yıkım yolları üzerine etkileri
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Dosyalar
Tarih
2019
Yazarlar
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Dergi ISSN
Cilt Başlığı
Yayıncı
Trakya Üniversitesi, Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Proteinlerin sentezinin, katlanmasının, taşınmasının ve yıkımlarının kontrolü proteostazın sürdürülmesi için şarttır. Hücre içi protein yıkımı ubikitin-proteazom sistemi ve otofaji sistemi tarafından gerçekleştirilmektedir. Mikrotübül-ilişkili protein 1A/1B-hafif zincir 3B protein otofaji markırlarından biridir ve fosfatidiletanolamin ile konjuge olarak mikrotübül-ilişkili protein 1A/1B-hafif zincir 3B-II olarak adlandırılan formda otofagazom yapısına girer. Isı şok proteini 70; proteinlerin katlanma, disagregasyonunda ve yıkımında rol oynayan önemli bir şaperondur. Literatürde karaciğerde akrilamidin ısı şok proteini 70 düzeylerine, ubiquitin-proteazom sistemine veya otofajiye etkisini araştıran bir çalışmaya rastlamadık. Bu çalışmada, insan kaynaklı hepatoma hücrelerinde akrilamidin protein yıkım yolları üzerine etkilerini araştırmayı amaçladık. Akrilamidin etkisinin araştırılması amacıyla HepG2 hücreleri 24 saat boyunca 10 1 , 10 2 , 10 3 ve 10 4 µM konsantrasyonlarda akrilamid ile inkübe edildi. Hücre canlılığı 3-(4,5Dimetil-2-tiazolil)-2,5-difenil-2H-tetrazolium bromür testi ile değerlendirildi. Isı şok proteini 70, ubikitinlenmiş protein ve mikrotübül-ilişkili protein 1A/1B-hafif zincir 3B-II protein düzeyleri western blot yöntemiyle ölçüldü. Akrilamid, 10 2 , 10 3 ve 10 4 µM konsantrasyonlarda hücre canlılığını anlamlı olarak azalttı. 10 4 µM akrilamid; ısı şok proteini 70, ubikitinlenmiş protein ve mikrotübül-ilişkili protein 1A/1B-hafif zincir 3B-II protein düzeylerinde anlamlı azalmaya yol açtı. Sonuç olarak, çalışmamız akrilamidin HepG2 hücrelerinde; hücre canlılığını azalttığını, ısı şok proteini 70, ubikitinlenmiş protein ve mikrotübül-ilişkili protein 1A/1Bhafif zincir 3B protein düzeylerini arttırdığını gösterdi. Bu çalışma ile insan kaynaklı hepatoma hücrelerinde akrilamidin özellikle yüksek konsantrasyonlarda protein yıkım yollarının çalışmasını bozduğu gösterilmiştir.
The control of synthesis, folding, transporting and degradation of proteins is essential for maintaining proteostasis. The degradation of intracellular proteins are performed by ubiquitin-proteasome system and autophagy. Microtubule-associated protein 1A/1B light chain 3B is an autophagy marker and enters to autophagasome structure by its conjugated form with phosphatidylethanolamine which is called as microtubule-associated protein 1A/1B light chain 3-II. Heat shock protein 70 is an important chaperone which plays role on the folding, dissagregation and degradation of proteins. In literature, we did not encounter any study which investigates the effect of acrylamide on heat shock protein 70 levels, ubiquitinproteasome system or autophagy in liver. In present study, we aimed to investigate the effect of acrylamide on protein degradation pathways in human-derived hepatoma cells. In order to investigate the effects of acrylamide, HepG2 cells were incubated with acrylamide at 10 2 , 10 3 ve 10 4 µM concentrations for 24 hours. Cell viability was evaluated by 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay. Heat shock protein 70, ubiquitinated protein and microtubule-associated protein 1A/1B light chain 3B protein levels were measured by western blotting. Acrylamide at 10 2 , 10 3 ve 10 4 µM concentrations significantly decreased cell viability. 10 4 µM acrylamide caused a significant increase on heat shock protein 70, ubiquitinated protein and microtubule-associated protein 1A/1B light chain 3B-II protein levels. As a result, our study showed that acrylamide decreases cell viability and increases heat shock protein 70, ubiquitinated protein and microtubule-associated protein 1A/1B lightchain 3B-II protein levels in HepG2 cells. With the present study, it has been shown that acrylamide, especially at high concentrations, disrupts protein degradation pathways in human-derived hepatoma cells.
The control of synthesis, folding, transporting and degradation of proteins is essential for maintaining proteostasis. The degradation of intracellular proteins are performed by ubiquitin-proteasome system and autophagy. Microtubule-associated protein 1A/1B light chain 3B is an autophagy marker and enters to autophagasome structure by its conjugated form with phosphatidylethanolamine which is called as microtubule-associated protein 1A/1B light chain 3-II. Heat shock protein 70 is an important chaperone which plays role on the folding, dissagregation and degradation of proteins. In literature, we did not encounter any study which investigates the effect of acrylamide on heat shock protein 70 levels, ubiquitinproteasome system or autophagy in liver. In present study, we aimed to investigate the effect of acrylamide on protein degradation pathways in human-derived hepatoma cells. In order to investigate the effects of acrylamide, HepG2 cells were incubated with acrylamide at 10 2 , 10 3 ve 10 4 µM concentrations for 24 hours. Cell viability was evaluated by 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay. Heat shock protein 70, ubiquitinated protein and microtubule-associated protein 1A/1B light chain 3B protein levels were measured by western blotting. Acrylamide at 10 2 , 10 3 ve 10 4 µM concentrations significantly decreased cell viability. 10 4 µM acrylamide caused a significant increase on heat shock protein 70, ubiquitinated protein and microtubule-associated protein 1A/1B light chain 3B-II protein levels. As a result, our study showed that acrylamide decreases cell viability and increases heat shock protein 70, ubiquitinated protein and microtubule-associated protein 1A/1B lightchain 3B-II protein levels in HepG2 cells. With the present study, it has been shown that acrylamide, especially at high concentrations, disrupts protein degradation pathways in human-derived hepatoma cells.
Açıklama
Anahtar Kelimeler
Akrilamid, HepG2, Isı Şok Proteini 70, Ubikitin-Proteazom Sistemi, Otofaji, Acrylamide, Heat Shock Protein 70, Ubiquitin-Proteasome System, Autophagy
