S-腺苷蛋氨酸通过抑制 Wnt2/β-Catenin 通路抑制视网膜母细胞瘤细胞 Y79 的增殖,诱导 Y79 细胞凋亡和细胞周期停滞。

IF 2.9 4区 医学 Q3 IMMUNOLOGY
Archivum Immunologiae et Therapiae Experimentalis Pub Date : 2024-10-04 eCollection Date: 2024-01-01 DOI:10.2478/aite-2024-0020
Mushi Liu, Youchaou Mobet, Hong Shen
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引用次数: 0

摘要

视网膜母细胞瘤是幼儿最常见的原发性眼内恶性肿瘤之一。化疗等传统治疗方法往往会带来严重的不良反应,如听力下降、认知障碍和视力减退。因此,迫切需要探索一种既有效又安全的新型治疗药物。众所周知,S-腺苷蛋氨酸(SAM)是一种天然化合物,在多种癌细胞系中具有抗增殖作用。然而,迄今为止,还没有研究调查过 SAM 对视网膜母细胞瘤细胞的影响及其潜在的作用机制。因此,本研究旨在调查亚麻酸对视网膜母细胞瘤细胞的影响,并探索其可能的作用机制,希望能为这种疾病的治疗提供新的见解。研究采用细胞计数试剂盒-8测定法确定了SAM的最佳浓度。使用 5-乙炔基-2'-脱氧尿苷细胞增殖试验评估了 SAM 对视网膜母细胞瘤增殖的影响。对 Y79 细胞进行苏木精和伊红染色,并用电子显微镜观察 SAM 诱导的任何形态变化。使用流式细胞术测量了 SAM 对视网膜母细胞瘤细胞周期的作用阶段及其凋亡效应。TUNEL 试验进一步证实了 SAM 对视网膜母细胞瘤的凋亡作用。通过 RT-PCR 检测了相关基因的差异表达。采用裸鼠体内皮下肿瘤形成和免疫组织化学方法验证了 SAM 对视网膜母细胞瘤相关表型的影响。通过 Western 印迹法研究 SAM 是否通过 Wnt2/β-catenin 通路调节视网膜母细胞瘤相关表型。结果表明,SAM能将视网膜母细胞瘤的细胞周期阻滞在G1期,通过Wnt2/β-catenin途径诱导视网膜母细胞瘤细胞凋亡,并影响其形态甚至超微结构。此外,体外和体内实验证明,SAM 对视网膜母细胞瘤有致癌作用。本研究在体外和体内验证了SAM是否能抑制视网膜母细胞瘤细胞Y7的增殖,通过抑制Wnt2/β-catenin通路诱导Y79细胞凋亡和细胞周期停滞,并影响视网膜母细胞瘤细胞Y79的形态和结构。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
S-Adenosylmethionine Inhibits the Proliferation of Retinoblastoma Cell Y79, Induces Apoptosis and Cell Cycle Arrest of Y79 Cells by Inhibiting the Wnt2/β-Catenin Pathway.

Retinoblastoma is one of the most common primary intraocular malignancies in young children. Traditional treatment methods such as chemotherapy often come with significant adverse effects, such as hearing loss, cognitive impairment, and vision loss. Therefore, there is an urgent need to explore a novel therapeutic drug that is both effective and safe. S-adenosylmethionine (SAM) is a natural compound known to exhibit anti-proliferative effects in various cancer cell lines. However, to date, no studies investigated the effects of SAM on retinoblastoma cells and its potential mechanisms of action. Therefore, this study aims to investigate the impact of SAM on retinoblastoma cells and explore its possible mechanisms of action, with the hope of providing new insights into the treatment of this disease. The optimal concentration of SAM was determined using the Cell Counting Kit-8 assay. The effect of SAM on retinoblastoma proliferation was assessed using the 5-ethynyl-2'-deoxyuridine cell proliferation assay. Y79 cells were subjected to hematoxylin and eosin stain and electron microscopy to observe any morphological changes induced by SAM. The stages of SAM's action on the retinoblastoma cell cycle and its apoptotic effects were measured using flow cytometry. The apoptotic effect of SAM on retinoblastoma was further confirmed using the TUNEL assay. Differential expression of related genes was detected through RT-PCR. In vivo subcutaneous tumor formation in nude mice and immunohistochemistry were employed to validate the effect of SAM on retinoblastoma-related phenotypes. Western blotting was conducted to investigate whether SAM modulated retinoblastoma-related phenotypes via the Wnt2/β-catenin pathway. SAM arrested the cell cycle of retinoblastoma at the G1 phase, induced apoptosis of retinoblastoma cells through the Wnt2/β-catenin pathway, and affected their morphology and even ultrastructure. In addition, in vitro and in vivo experiments demonstrated that SAM had an oncogenic effect on retinoblastoma. In this study, we verify in vitro and in vivo whether SAM inhibits the proliferation of retinoblastoma cell Y7, induces apoptosis and cell cycle arrest of Y79 cells by inhibiting the Wnt2/β-catenin pathway, and affects the morphology and structure of retinoblastoma cell Y79.

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来源期刊
CiteScore
5.90
自引率
0.00%
发文量
26
审稿时长
>12 weeks
期刊介绍: Archivum Immunologiae et Therapiae Experimentalis (AITE), founded in 1953 by Ludwik Hirszfeld, is a bimonthly, multidisciplinary journal. It publishes reviews and full original papers dealing with immunology, experimental therapy, immunogenetics, transplantation, microbiology, immunochemistry and ethics in science.
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