摘要
目的:检测环状RNA CircCrebrf在细胞复制性衰老模型中的表达水平及亚细胞定位情况。方法:基于小鼠胚胎成纤维细胞,连续传代至P9代,从而建立细胞复制性衰老模型;利用细胞衰老β-半乳糖苷酶(SA-β-gal)染色及Western blot检测p16蛋白的表达验证细胞衰老表型;利用qPCR检测CircCrebrf在衰老细胞中的表达;核质分离实验分析CircCrebrf亚细胞定位。结果:CircCrebrf在P9代衰老细胞中的表达水平明显上升,并主要定位于细胞质中,提示其可能在细胞衰老进程发挥作用。结论:环状RNA CircCrebrf的表达与细胞复制性衰老进程密切相关。
Objective:To evaluste the expression level and subcellular location of Circular RNA CircCrebrf in a replicative cellular senescence model.Methods:This study establish a replicative cellular senescence model utilizing serial passage to P9 generation of MEF cells;SA-β-gal staining and examination of P16 protein levels by Western blot are performed to verify the senescence phenotype of cells;qPCR is carried out to determine the CircCrebrf expression levels in senescent cells;nuclear-cytoplasmic separation experiment is perform to analyze the subcellular location of CircCrebrf.Results:The results uncover that CircCrebrf is up-regulated in senescent P9 generation of MEF cells,and is localized in cytoplasm,which indicate the potential role of CircCrebrf in cellular senescence.Conclusion:The expression of circular RNA CircCrebrf is closely related to the process of cell replicative senescence.
作者
徐舜
雷仪婷
沈佳恩
郑晓丽
吴赟
XU Shun;LEI Yiting;SHEN Jiaen;ZHENG Xiaoli;WU Yun(Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnosis,Guangdong Medical University,Dongguan 523808,China;Institute of Aging,Guangdong Medical University,Dongguan 523808,China)
出处
《生物化工》
2025年第4期141-144,共4页
Biological Chemical Engineering
基金
广东大学生科技创新培育专项资金(攀登计划专项资金)(pdjh2021b0226)
大学生创新创业训练计划项目(GDMU2023167,202310571038,ZZDC002)。
作者简介
徐舜(1984-),男,浙江诸暨人,博士,副研究员,研究方向为衰老分子生物学;通信作者:吴赟(1987-),男,江西泰和人,博士,助理研究员,研究方向为衰老分子生物学。E-mail:wuyun@gdmu.edu.cn。
