【作者】 黄江；

【导师】 祝新根；

【作者基本信息】 南昌大学 ， 临床医学博士（专业学位）， 2025， 博士

【摘要】 背景:肿瘤蛋白p53诱导核蛋白2(tumor protein p53-inducible nuclear protein2,TP53INP2)在多种肿瘤中被发现表达失调,并能通过分子机制调节肿瘤恶性进展。然而,TP53INP2在胶质瘤中尚无报道,鉴于其在肿瘤中的重要作用,亟需探究其在胶质瘤中的作用。方法:采用RNA干扰技术靶向沉默TP53INP2基因表达,将si-NC或si-TP53INP2-1/2与U251细胞共培养,转染后的细胞进行体外实验。采用实时定量聚合酶链反应法检测体外TP53INP2 m RNA的表达。Western blotting检测TP53INP2、上皮-间充质转化(epithelial-to-mesenchymal transition,EMT)相关蛋白和NF-κB/Nrf2/HO-1通路相关蛋白水平。Annexin V-FITC/PI双标记法进行细胞凋亡检测,采用2’,7’-二氯二氢荧光素二乙酸酯(DCFH-DA)荧光探针法,结合流式细胞术定量分析细胞内ROS水平。本研究通过多维度技术(CCK-8细胞活力检测、Ed U DNA增殖标记分析及克隆形成实验)系统评估U251胶质瘤细胞的增殖能力,同时进行transwell检测以评估其迁移和侵袭能力。流式细胞仪检测凋亡细胞及活性氧含量。采用酶联免疫吸附法测定超氧化物歧化酶和谷胱甘肽过氧化物酶水平。此外,建立了小鼠异种肿瘤移植模型进行肿瘤细胞异种移植研究。结果:胶质瘤细胞中TP53INP2的表达明显增高。TP53INP2缺失显著抑制肿瘤生长、转移、EMT过程和氧化应激。此外,被沉默的TP53INP2被证实可以抑制NF-κB的表达,促进Nrf2/HO-1通路的激活,同时降低了免疫细胞凋亡率并增加了其数量,从而抑制胶质瘤的生长。结论:TP53INP2/NF-κB/Nrf2/HO-1信号通路调控胶质瘤生长的病理过程,为治疗胶质瘤提供了新的途径和理论依据。更多还原

【Abstract】 Background:The function of tumor protein p53-inducible nuclear protein 2(TP53INP2)in numerous cancers has been elucidate clearly;however,its role across the development of gliomas remain largely unexplored.Methods:Quantitative real-time polymerase chain reaction was employed to determine TP53INP2 m RNA expression in vitro.Western blotting was subjected to detect TP53INP2,epithelial-to-mesenchymal transition(EMT)-related and NF-κB/Nrf2/HO-1 pathway related protein levels.The proliferative potentials of glioma cells were assessed by 5-ethynyl-2’-deoxyuridine,colony formation,and cell counting kit-8 assays,while performing transwell assays to evaluate the migratory and invasive capacities.Apoptotic cells and reactive oxygen species contents were analyzed using flow cytometer.Enzyme-linked immunosorbent assay was performed to measure superoxide dismutase and glutathione peroxidase levels.Additionally,tumor xenograft model in mouse was also conducted.Results:Pronounced higher expression of TP53INP2 was observed in glioma cells.TP53INP2 depletion showed a significant inhibition in tumor growth,metastasis,EMT process and oxidative stress.Moreover,the silenced TP53INP2 was confirmed to hamper the activation of NF-κB and promote the activation of Nrf2/HO-1 pathway,and decrease apoptosis rate and increase the number of immune cells.Conclusions:The above-mentioned results highlight the therapeutic potential of TP53INP2/NF-κB/Nrf2/HO-1 axis in gliomas.更多还原