| hnRNPU过表达抑制PIM1诱导的细胞衰老的机制研究 |
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| 引用本文： | 肖雅雯,阳检明.hnRNPU过表达抑制PIM1诱导的细胞衰老的机制研究[J].天津医科大学学报,2023,0(2):137-141，169. |
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| 作者姓名： | 肖雅雯 阳检明 |
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| 作者单位： | 天津医科大学基础医学院免疫学系，天津300070 |
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| 摘 要： | 目的:探讨人前病毒整合位点1（PIM1）诱导细胞衰老的分子机制。方法:构建过表达PIM1的2BS细胞系，通过Western印迹法和衰老相关β-半乳糖苷酶染色实验测定过表达PIM1是否诱导细胞衰老。免疫沉淀联合质谱分析测定PIM1是否能有效免疫沉淀核异质核糖核蛋白U（hnRNPU）蛋白。运用Real-timePCR和Western印迹法测定PIM1是否影响hnRNPUmRNA和蛋白表达水平。构建同时过表达PIM1和hnRNPU的2BS细胞系，运用Western印迹法和衰老相关β-半乳糖苷酶染色实验检测hnNRPU过表达对PIM1诱导的细胞衰老的影响。结果:与空载对照组相比，过表达PIM1组中衰老信号通路关键基因p53、p21和p16的表达显著增加（p53，t=4.36，P<0.05；p21，t=3.814，P<0.05；p16，t=4.72，P<0.01），衰老细胞的比例增加（t=6.831，P<0.01）。免疫沉淀联合质谱分析结果显示PIM1能有效免疫沉淀hnRNPU蛋白。PIM1过表达对hnRNPU的mRNA表达水平没有影响（t=0.295，P=0.783），但是能抑制hnRNPU蛋白的表达（t=33.85，P<0.001）。与只过表达PIM1细胞相比，同时过表达PIM1和hnRNPU细胞，衰老信号通路关键基因p53、p21和p16的表达下降（p53，t=15.317，P<0.001；p21，t=8.012，P<0.01；p16，t=14.08，P<0.001），衰老细胞的比例降低（t=10.38，P<0.01）。结论:hnRNPU过表达抑制PIM1诱导的细胞衰老。
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| 关 键 词： | 人前病毒整合位点1 核异质核糖核蛋白U 细胞衰老 |
Study of the mechanism of hnRNPU overexpression inhibiting PIM1-induced cellular senescence |
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| XIAO Ya-wen,YANG Jian-ming.Study of the mechanism of hnRNPU overexpression inhibiting PIM1-induced cellular senescence[J].Journal of Tianjin Medical University,2023,0(2):137-141，169. |
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| Authors: | XIAO Ya-wen YANG Jian-ming |
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| Affiliation: | Department of Immunology，School of Basic Medical Sciences，Tianjin Medical University，Tianjin 300070，China |
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| Abstract: | Objective:To investigate the molecular mechanism of proviral integration site 1（PIM1）-induced cellular senescence. Methods:The human embryonic lung fibroblasts 2BS cell overexpressing PIM1 was constructed. Western blotting and senescence-related β-galactosidase staining experiments was used to determine whether overexpression of PIM1 induced cellular senescence. Immunoprecipitation combined with mass spectrometry was used to determine whether PIM1 could effectively immunoprecipitate nuclear heterogeneous ribonucleoprotein U（hnRNPU）protein. The hnRNPU mRNA and protein levels were evaluated by real-time PCR and Western blotting. The 2BS cell overexpressing PIM1 and hnRNPU was constructed and the effect of hnNRPU overexpression on PIM1-induced cellular senescence was detected by Western blotting and senescence-related β-galactosidase staining assay. Results:Compared with the control group，the expression of p53，p21，and p16，key genes in regulating cellular senescence were significantly increased（p53，t=4.36，P<0.05；p21，t=3.814，P<0.05；p16，t=4.72，P<0.01）and the proportion of senescent cells increased（t=6.831， P<0.01）in the PIM1 overexpression group. Immunoprecipitation combined with mass spectrometry showed that PIM1 could effectively immunoprecipitate hnRNPU protein. PIM1 overexpression had no effect on hnRNPU mRNA expression （t=0.295，P=0.783），but downregulated the hnRNPU protein expression（t=33.85，P<0.001）. Compared with PIM1 group，the expression of p53，p21，and p16，key genes in regulating cellular senescence，were decreased in the overexpression of hnRNPU and PIM1 group（p53，t=15.317，P<0.001;p21，t=8.012，P<0.01; p16，t=14.08，P<0.001）. Compared with PIM1 group，the overexpression of hnRNPU and PIM1 group showed lower proportion of senescent cells（t=10.38，P<0.01）. Conclusion:Overexpression of hnRNPU inhibits PIM1-induced cellular senescence. |
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| Keywords: | proviral integration site 1 heterogeneous nuclear ribonucleoprotein U cell senescence |
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