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.
Studies on the Potential Role of DEAD-box Helicase 41 in DNA Damage Response
To investigate the impact of DEAD-box Helicase 41(DDX41)knockdown on DNA Damage Re-pair(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 experi-ments.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 mainte-nance,offering insights for cancer prevention and treatment strategies.
DNA Damage RepairDEAD-box Helicase 41Ubiquitin-Specific-Processing Protease 11
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