PSMC4表达下调促进MEF细胞衰老  

Down-regulation of PSMC4 expression promotes MEF cell senescence

作　　者：骆诗凌 谢健鸿 吴世裕 曾芷彤 刘新光 袁源 LUO Shiling;XIE Jianhong;WU Shiyu;ZENG Zhitong;LIU Xinguang;YUAN Yuan(Guandong Provincial Key Laboratory of Medical Molecular Diagnostics,Dongguan 523808;Institute of Aging Research,Guangdong Medical University,Dongguan 523808;Institute of Biochemistry&Molecular Biology,Guangdong Medical University,Dongguan 523808,China)

机构地区：[1]广东省医学分子诊断重点实验室,广东东莞523808 [2]广东医科大学衰老研究所,广东东莞523808 [3]广东医科大学生物化学与分子生物学研究所,广东东莞523808

出　　处：《生物技术》2024年第5期543-547,共5页Biotechnology

基　　金：国家自然科学基金面上项目(81971309,81671399);广东医科大学博士学位人员科研启动基金项目(GDMUB2023005);广东医科大学大学生创新实验项目(2020ZYDB002,2020ZZDB003);广东省大学生创新创业训练计划项目(S202210571066)。

摘　　要：[目的]探讨PSMC4表达下调对小鼠胚胎成纤维(Mouse Embryonic Fibroblast,MEF)细胞衰老的影响。[方法]利用小干扰RNA(siRNA)敲低PSMC4,荧光定量PCR和Western Blotting验证siRNA的敲低效果;通过β-半乳糖苷酶染色检测细胞衰老情况;通过EdU检测细胞增殖;通过酶联免疫吸附法检测26S蛋白酶体活性。[结果]下调PSMC4后β-半乳糖苷酶染色阳性细胞数量明显增加,EdU细胞增殖率降低约2.7倍(P<0.01),26S蛋白酶体活性降低约1.6倍(P<0.01)。[结论]PSMC4表达下调通过降低26S蛋白酶体活性抑制MEF细胞增殖,进而促进MEF细胞衰老。[Objective]To investigate the impact of down-regulated PSMC4 expression on Mouse Embryonic Fibroblast(MEF)cell senescence.[Method]PSMC4 was targeted for knockdown using small interfering RNA(siRNA),and the knockdown efficiency was validated by quantitative PCR and Western Blotting.Cellular senescence was detected byβ-galactosidase.Cellular proliferation was evaluated using EdU.26S proteasome activity was determined via enzyme-linked immunosorbent assay.[Result]Down-regulation of PSMC4 significantly increased the number ofβ-galactosidase staining positive cells,decreased the proliferation rate of EdU cells by about 2.7-fold(P<0.01),and decreased the 26S proteasome activity by about 1.6-fold(P<0.01).[Conclusion]The down-regulation of PSMC4 expression hindered MEF cell proliferation by diminishing 26S proteasome activity,consequently promoting MEF cell senescence.

关 键 词：蛋白酶体 小鼠胚胎成纤维细胞 PSMC4 SIRNA 细胞衰老 细胞增殖 

分 类 号：Q291[生物学—细胞生物学]