SH-SY5Y nöroblastoma hücresinde soğuk stresin transkriptomik etkilerinin araştırılması
Küçük Resim Yok
Tarih
2026
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
İnönü Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Amaç: Bu çalışmanın amacı soğuk stresinin nöroblastoma hücreleri üzerine etkilerinin transkriptomik ve protein seviyeleri düzeyinde araştırmaktır. Materyal ve Metot: Çalışmada SH-SY5Y nöroblastoma hücre hattı kullanıldı. Soğuk stres modelinde kullanılan hücreler 37°C (Kontrol), 31°C ve 25°C sıcaklıklarında ve 3, 6, 12, 24 saat sürelerinde inkübe edildi. Her sürenin sonunda RNA sekans verileri ve gen ekpresyonu için RNA izolasyonu, western blot için protein ekspresyonu, apoptoz düzeyleri için Annexin-V FITC yöntemi, hücre döngüsü için PI analizi yapıldı. İstatistiksel veriler için tek yönlü ANOVA testi uygulanıp sonuçlar değerlendirildi. Bulgular: SH-SY5Y hücrelerinde düşük sıcaklığın etkileri değerlendirildi ve özellikle 25 °C'de hücre morfolojisinin belirgin şekilde bozulduğu görüldü. RNA-seq analizleri, soğuk stresinin özellikle 31 °C 3 saat ve 25 °C 24 saat koşullarında anlamlı gen baskılanmasına neden olduğu saptandı. qPCR ve Western blot seviyeleri ATF6, HSP90 ve LIN28A'nın bazı koşullarda arttığını, CARHSP1, TP53, UBA52 ve XBP1'in ise çoğunlukla azaldığı saptandı. 25 °C'de inkübasyon süresi uzadıkça apoptoz oranlarının arttığı ve hücre döngüsü fazlarında belirgin değişimler meydana geldiği belirlendi. Sonuç: Çalışmamız da düşük sıcaklık stresinin SH-SY5Y hücrelerinde morfoloji, gen/protein ifadesi, apoptoz ve hücre döngüsü üzerinde belirgin etkiler oluşturduğunu özellikle 25 °C'de hem hücresel bütünlük bozulmuş hem de genetik yanıtlar güçlü şekilde değişmiş olup soğuk stresinin SH-SY5Y hücrelerinde hem moleküler hem de fonksiyonel düzeyde yeniden programladığını ortaya koymaktadır. Anahtar Kelimeler: Kanser, Mikroçevre, Soğuk stresi, SH-SY5Y
Aim: The aim of this study is to investigate the effects of cold stress on neuroblastoma cells at the transcriptomic and protein levels. Materials and Methods: The SH-SY5Y neuroblastoma cell line was used in this study. Cells were incubated at 37°C (Control), 31°C, and 25°C for 3, 6, 12, and 24 hours under a cold stress model. At the end of each time point, RNA isolation was performed for sequencing and gene expression analysis, protein expression was evaluated by Western blot, apoptosis levels were measured using the Annexin-V FITC method, and cell cycle distribution was assessed using PI analysis. Statistical evaluation was conducted using one-way ANOVA. Results: The effects of low temperature on SH-SY5Y cells were evaluated, and cell morphology was found to be markedly impaired, particularly at 25 °C. RNA-seq analyses revealed that cold stress led to large-scale, significant gene suppression, especially under the conditions of 31 °C for 3 hours and 25 °C for 24 hours. qPCR and Western blot results showed that ATF6, HSP90, and LIN28A increased under certain conditions, whereas CARHSP1, TP53, UBA52, and XBP1 mostly decreased. It was determined that the rate of apoptosis increased and significant alterations occurred in cell cycle phases as the incubation time at 25 °C was extended. Conclusion: This study demonstrates that cold stress induces notable changes in morphology, gene/protein expression, apoptosis, and cell cycle dynamics in SH-SY5Y cells. Particularly at 25°C, both cellular integrity and genetic responses were markedly affected, indicating that cold stress leads to molecular and functional reprogramming in SH-SY5Y cells. Key Words: Cancer, Microenvironment, Cold stress, SH-SY5Y
Aim: The aim of this study is to investigate the effects of cold stress on neuroblastoma cells at the transcriptomic and protein levels. Materials and Methods: The SH-SY5Y neuroblastoma cell line was used in this study. Cells were incubated at 37°C (Control), 31°C, and 25°C for 3, 6, 12, and 24 hours under a cold stress model. At the end of each time point, RNA isolation was performed for sequencing and gene expression analysis, protein expression was evaluated by Western blot, apoptosis levels were measured using the Annexin-V FITC method, and cell cycle distribution was assessed using PI analysis. Statistical evaluation was conducted using one-way ANOVA. Results: The effects of low temperature on SH-SY5Y cells were evaluated, and cell morphology was found to be markedly impaired, particularly at 25 °C. RNA-seq analyses revealed that cold stress led to large-scale, significant gene suppression, especially under the conditions of 31 °C for 3 hours and 25 °C for 24 hours. qPCR and Western blot results showed that ATF6, HSP90, and LIN28A increased under certain conditions, whereas CARHSP1, TP53, UBA52, and XBP1 mostly decreased. It was determined that the rate of apoptosis increased and significant alterations occurred in cell cycle phases as the incubation time at 25 °C was extended. Conclusion: This study demonstrates that cold stress induces notable changes in morphology, gene/protein expression, apoptosis, and cell cycle dynamics in SH-SY5Y cells. Particularly at 25°C, both cellular integrity and genetic responses were markedly affected, indicating that cold stress leads to molecular and functional reprogramming in SH-SY5Y cells. Key Words: Cancer, Microenvironment, Cold stress, SH-SY5Y
Açıklama
Anahtar Kelimeler
Tıbbi Biyoloji, Medical Biology
