转录因子YAP1在X线诱导的HaCaT细胞损伤中的作用和作用机制  

Study on the role and mechanism of transcription factor YAP1 in X-ray-induced damage of HaCaT cells

作　　者：黄腾 于大海 王万霞 鹿红 HUANG Teng;YU Dahai;WANG Wanxia;LU Hong(Department of Radiotherapy,Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China)

机构地区：[1]南京中医药大学附属医院放疗科,江苏南京210029

出　　处：《生物加工过程》2025年第2期207-211,共5页Chinese Journal of Bioprocess Engineering

基　　金：国家自然科学基金青年项目(82405508);江苏省中医药管理局科技项目(YB2020009、YB201815)。

摘　　要：放射性皮肤损伤是放射治疗过程中常见的并发症。利用人永生化表皮细胞(HaCaT),采用X线体外照射,构建放射性皮肤损伤细胞模型,并采用四甲基偶氮唑盐(MTT)比色法,5乙炔基2’脱氧尿苷(EDU)增殖实验和单克隆实验证实:X线能够抑制HaCaT细胞的活性和增殖能力。通过2’,7’二氯荧光素二乙酸酯(DCFH DA)和二氢乙锭(DHE)探针检测X线对细胞内活性氧(ROS)水平的影响,并通过定量聚合酶链反应(qPCR),Western blotting和免疫荧光实验探讨X线抑制细胞增殖的机制。结果显示:X线能够抑制HaCaT细胞活力和增殖能力,诱导细胞内ROS的增加,下调细胞周期蛋白D1(CyclinD1)和细胞周期蛋白E1(CyclinE1)的表达。进一步的机制研究显示:X线能够抑制转录因子Yes相关蛋白1(YAP1)的表达和核转位,可能是导致细胞增殖能力降低的重要原因。本研究揭示了X线造成HaCaT细胞损伤的分子机制,为预防和治疗放射性皮肤损伤提供了分子靶点。Radiation-induced skin injury is a common complication of radiotherapy.In this study,using HaCaT cells,a cell model of radiation-induced skin injury was developed through in vitro X-ray irradiation.The inhibitory effects of X-rays on the viability and proliferation capacity of HaCaT cells were validated through the MTT colorimetric assay,EDU proliferation assay,and colony formation assay.DCFH-DA and DHE probes were used to assess the effects of X-ray on intracellular reactive oxygen species(ROS)levels.The mechanism of X-ray-induced inhibition of cell proliferation by RT-qPCR,Western-blotting,and immunofluorescence assay.The results showed that X-ray inhibited the viability and proliferation capacity of HaCaT cells,induced an increase in intracellular ROS levels,and downregulated the expression of CyclinD1 and CyclinE1.Moreover,X-ray inhibited the expression and nuclear translocation of the transcription factor YAP1,which may be an important reason for the reduced cell proliferation capacity.This study revealed the molecular mechanism of X-ray-induced damage of HaCaT cells and identified a molecular target for the prevention and treatment of radiation-induced skin injury.

关 键 词：放射损伤 HACAT细胞 YAP1 细胞增殖 氧化应激 

分 类 号：R965[医药卫生—药理学]