摘要
目的探讨泛素结合酶E2变体蛋白(UBE2V1)对MEFs细胞衰老的影响。方法Western blot检测UBE2V1在年轻(2月龄)和年老小鼠(24月龄)各组织中的表达差异,以荧光定量PCR及Western blot检测UBE2V1在年轻和衰老MEFs细胞中表达的差异,荧光定量PCR和Western blot验证Ube2v1基因的siRNA沉默效果,β-半乳糖苷酶染色检测Ube2v1表达下调对MEFs细胞衰老的影响,Edu和Alarmar Blue检测Ube2v1基因沉默后MEFs细胞增殖能力的变化。结果UBE2V1在衰老小鼠的心脏、肝脏、肾脏和脑组织中表达下调,同时在衰老的MEFs细胞中也明显表达下调,在年轻细胞中沉默Ube2v1基因后β-半乳糖苷酶染色结果显示衰老的细胞数量明显增加,且Edu和Alarmar Blue检测发现Ube2v1基因沉默后MEFs细胞增殖能力显著下降。结论沉默Ube2v1可通过抑制MEFs细胞增殖,从而促进细胞衰老。
Objective To investigate the effect of ubiquitin-binding enzyme E2 variant(UBE2V1)on cellular senescence of MEFs.Methods Western blot was used to detect the difference of UBE2V1 expression in the tissues of young(2 months of age)and old(24 months of age)mice.Quantitative PCR and Western blot were used to detect the difference of UBE2V1 expression in young and senile MEFs cells.Quantitative PCR and Western blot were used to verify the siRNA silencing effect of Ube2v1 gene.β-galactosidase staining was used to detect the influence of downregulation of Ube2v1 expression on aging of MEFs cells.Edu and Alarmar Blue were used to detect the changes of proliferation ability of MEFs cells after Ube2v1 gene silencing.Results Ube2v1 was down-regulated in the heart,liver,kidney and brain tissues of aging mice,and also downregulated in aging MEFs cells.β-galactosidase staining results showed that the number of aging cells increased significantly after Ube2v1 gene was silenced in young cells.Edu and Alarmar Blue tests showed that the proliferation ability of MEFs cells decreased significantly after Ube2v1 gene silencing.Conclusion Silencing Ube2v1 inhibits MEFs cell proliferation and thus promote cell senescence.
作者
徐舜
骆诗凌
谢健鸿
刘新光
袁源
XU Shun;LUO Shi-ling;XIE Jian-hong;LIU Xin-guang;YUAN Yuan(Guangdong Provincial KeyLaboratory of Medical Molecular Diagnostics,Guangdong Medical University,Dongguan 523808,China;Instituteof Aging Research,Guangdong Medical University,Dongguan 523808,China;Institute of Biochemistry&MolecularBiology,Guangdong Medical University,Dongguan 523808,China)
出处
《广东医科大学学报》
2023年第3期255-258,共4页
Journal of Guangdong Medical University
基金
国家自然科学基金(81671399)
广东省医学科研基金(A2020173)
广东医科大学科研基金面上培育项目(GDMUM201803)
广东省大学生创新创业训练计划项目(201810571105,S202110571102,S202210571066)
广东医科大学大学生创新实验项目(2020ZYDB002,2020ZZDB003)
广东医科大学科研基金自然科学类重点培育项目(GDMUZ2020003)。
