PSMC4表达下调促进MEF细胞衰老

Down-regulation of PSMC4 expression promotes MEF cell senescence

[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.