Investigation of the transcriptomic effects of cold stress in SH-SY5Y neuroblastoma cells

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