目的 对在模式生物斑马鱼中紫外线(UV)和依托泊苷(Epotoside,ETO)分别诱导的DNA损伤条件下Ypel3与p53信号通路的应答反应进行研究.方法 利用CRISPR/Cas9基因编辑技术,在模式生物斑马鱼中构建ypel3基因敲除突变体,通过UV辐射及抗癌药物ETO处理斑马鱼胚胎,结合实时荧光定量PCR技术,探究在物、化异常产生的DNA损伤环境中,Ypel3与p53信号通路间的互作关系.结果 在受精后24 h(hours post fertilization,hpf),UV辐射诱导DNA损伤引起野生型斑马鱼胚胎p53转录水平上调,ypel3也随之显著上调;与野生型胚胎相比,ypel3突变体受UV辐射后p53转录水平上调更为显著.然而,在ETO诱导的斑马鱼DNA损伤模型中,与野生型胚胎相比,ypel3突变对p53转录水平的调控却略有下调.结论 斑马鱼中验证了 ypel3是p53信号通路的靶基因,在UV物理损伤及ETO化学物质诱导的DNA损伤模型中,Ypel3对p53转录水平发挥不同的作用,提示Ypel3对抗肿瘤免疫反应有重要影响.聚焦肿瘤相关DNA损伤,构建UV诱导的皮肤癌相关药物筛选模型,为抗肿瘤和抗肿瘤免疫反应相关研究提供新思路.
Exploration the interaction between Ypel3 and p53 in zebrafish DNA lesion models
Objective To study the response of Ype13(Yippee-like 3)and the p53(tumor protein 53)signaling pathway to DNA damage which inducing by ultraviolet(UV)radiation and Epotoside(ETO).Methods Using CRISPR/Cas9 gene editing technology,we constructed ypel3 knockout mutant in the model organism zebrafish,and treated zebrafish embryos with UV radiation and ETO.We used real-time fluorescence quantitative PCR to explore the interaction between Ype13 and p53 pathway in DNA lesion models.Results At 24 hours post-fertilization(hpf),UV radiation-induced DNA lesion caused p53 transcription level to be upregulated in wild type zebrafish embryos,and ypel3 was also significantly upregulated.Compared with wild type embryos,the ypel3 mutant showed a more significant upregulation of p53 transcription level after UV radiation.However,in the zebrafish DNA damage model induced by ETO,the ypel3 MZ mutants showed a slight downregulation in the regulation of p53 transcription levels compared with wide type embryos.Conclusion This study verified that ypel3 was the target gene of the p53 signaling pathway in zebrafish,and in the UV physical damage and ETO chemical damage-induced DNA lesion models,Ypel3 played different roles in regulating p53 transcription levels,suggesting that Ypel3 has an important effect on anti-tumor immune response.This study focused on tumor-related DNA lesion and constructed a UV-induced skin cancer related drug screening model,providing new ideas for the study of anti-tumor immune response.
ypel3zebrafishmodel of screening anti-tumor drugsp53
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维普
