DNA损伤应答中DEAD-box Helicase 41的潜在作用研究  

Studies on the Potential Role of DEAD-box Helicase 41 in DNA Damage Response

作　　者：王心宇 朱敏[1] WANG Xin-yu;ZHU Min(Institute for Translation Medicine,Qingdao University,Qingdao 266021,China)

机构地区：[1]青岛大学转化医学研究院,青岛266021

出　　处：《青岛大学学报(自然科学版)》2024年第4期8-12,18,共6页Journal of Qingdao University(Natural Science Edition)

基　　金：国家自然科学基金(批准号:32201061)资助;山东省自然科学基金(批准号:ZR2021QC083)资助。

摘　　要：为探究死盒解旋酶41(DEAD-box Helicase 41,DDX41)的敲低如何影响DNA损伤应答(DNA Damage Repair,DDR),慢病毒筛选敲低DDX41(shDDX41)的HeLa细胞,转染质粒FLAG-USP11、GFP-DDX41至HEK-293T细胞,设置细胞空白对照组(NC细胞)和依托泊苷加药组,基于Western blot、免疫荧光染色以及染色体分离实验,确定shDDX41细胞DNA损伤情况及其DDR情况。实验结果显示,shDDX41细胞自发性DNA损伤高于NC细胞,细胞染色体畸形比例明显增加;shDDX41细胞过表达去泛素化酶(Ubiquitin-Specific-Processing Protease 11,USP11)后染色体畸形比例明显降低。shDDX41会募集肿瘤蛋白p53结合蛋白及组蛋白H2AX影响DDR过程。DDX41与USP11的相互作用受DNA损伤调控。这表明,DDX41可参与DDR,维护基因组稳定性,可为癌症预防治疗提供参考。To investigate the impact of DEAD-box Helicase 41(DDX41)knockdown on DNA Damage Repair(DDR),HeLa cells were screened using lentiviral vectors for DDX41 knockdown(shDDX41),and HEK-293T cells were transfected with FLAG-USP11 and GFP-DDX41 plasmids.A blank control group(NC cells)and an etoposide-treated group were established to assess DNA damage and DDR in shDDX41 cells through Western blot analysis,immunofluorescence staining,and chromosome segregation experiments.The results demonstrate that spontaneous DNA damage is higher in shDDX41 cells compared to NC cells,with a significant increase in the proportion of cellular chromosomal aberrations.Overexpression of Ubiquitin-Specific-Processing Protease 11(USP11)in shDDX41 cells notably reduce the proportion of chromosomal aberrations.shDDX41 is able to recruit tumor protein p53 binding protein 1 and histone H2AX to the DDR process.Furthermore,the interaction between DDX41 and USP11 is found to be DNA damage-dependent.These findings suggest that DDX41 plays a role in DDR and genomic stability maintenance,offering insights for cancer prevention and treatment strategies.

关 键 词：DNA损伤应答 死盒解旋酶41 去泛素化酶 

分 类 号：Q527[生物学—生物化学]