Kurt Serap, Karagur Ege Riza, Kavak Deniz Evrim, Kizildag Sefa
Department of Medical Biology and Genetics, Institute of Health Sciences, Dokuz Eylul University, İzmir, Turkey.
Department of Medical Genetics, Faculty of Medicine, Manisa Celal Bayar University, Manisa, Turkey.
Mol Biol Rep. 2025 Jun 6;52(1):557. doi: 10.1007/s11033-025-10628-7.
Alzheimer's disease (AD) is a diverse neurodegenerative disorder that is defined by impairments in cognitive function.
This study seeks to examine the apoptotic function of long non-coding RNA linc00968 in amyloid beta 25-35 neurotoxic SH-SY5Y cells.
Fragments of Aβ25-35 were administered to SH-SY5Y cells in order to simulate the neurotoxicity associated with Alzheimer's disease. Cell viability was assessed using the MTT test. Using quantitative real-time polymerase chain reaction, the expression levels of linc00968 were examined in treated and untreated (control) cells exposed to Aβ25-35. Subsequently, linc00968 was transfected with siRNA into Aβ25-35-treated SH-SY5Y cells to silence its expression and confirmed by qRT-PCR. The impact of BCL-2, BAX, CYT-C, and the internal control gene GAPDH on apoptotic pathways was examined using quantitative real-time polymerase chain reaction (qRT-PCR). To assess the post-transcriptional effects, the levels of Bcl-2, Bax, Cyt-c, and Beta-actin were analyzed using western blotting. The expression of Linc00968 was observed to be elevated in Aβ 25-35 stimulated SH-SY5Y cells. Aβ25-35-mediated toxicity in SH-SY5Y cells led to a reduction in the expression of the anti-apoptotic gene BCL-2, while the expressions of the pro-apoptotic genes BAX and CYT-C were found to be elevated. Suppression of linc00968 was observed to reverse apoptosis in Aβ25-35-induced SH-SY5Y cells. At the protein level, silencing linc00968 with siRNA resulted in elevated levels of the anti-apoptotic protein Bcl-2 and decreased levels of the pro-apoptotic proteins Bax and Cyt-c.
Our results show that Aβ25-35-induced SH-SY5Y cells exhibit elevated expression of linc00968. By controlling several apoptosis-related indicators, we proposed that inhibition of linc00968 can shield Aβ25-35 produced neurotoxicity in SH-SY5Y cells.
阿尔茨海默病（AD）是一种多样的神经退行性疾病，由认知功能障碍所定义。
本研究旨在检测长链非编码RNA linc00968在淀粉样β蛋白25 - 35神经毒性的SH - SY5Y细胞中的凋亡功能。
将Aβ25 - 35片段作用于SH - SY5Y细胞，以模拟与阿尔茨海默病相关的神经毒性。使用MTT试验评估细胞活力。采用定量实时聚合酶链反应检测在暴露于Aβ25 - 35的处理和未处理（对照）细胞中linc00968的表达水平。随后，将linc00968用小干扰RNA转染至经Aβ25 - 35处理的SH - SY5Y细胞中以沉默其表达，并通过定量逆转录 - 聚合酶链反应进行确认。使用定量实时聚合酶链反应（qRT - PCR）检测BCL - 2、BAX、CYT - C和内参基因GAPDH对凋亡途径的影响。为评估转录后效应，使用蛋白质印迹法分析Bcl - 2、Bax、Cyt - c和β - 肌动蛋白的水平。观察到在Aβ25 - 35刺激的SH - SY5Y细胞中Linc00968的表达升高。Aβ25 - 35介导的SH - SY5Y细胞毒性导致抗凋亡基因BCL - 2的表达降低，而促凋亡基因BAX和CYT - C的表达升高。观察到抑制linc00968可逆转Aβ25 - 35诱导的SH - SY5Y细胞凋亡。在蛋白质水平，用小干扰RNA沉默linc00968导致抗凋亡蛋白Bcl - 2水平升高，促凋亡蛋白Bax和Cyt - c水平降低。
我们的结果表明，Aβ25 - 35诱导的SH - SY5Y细胞表现出linc00968表达升高。通过控制多个凋亡相关指标，我们提出抑制linc00968可保护SH - SY5Y细胞免受Aβ25 - 35产生的神经毒性。
