SOX5介导的DNA损伤修复在下咽癌顺铂耐药性中的作用及其机制研究  

Role of SOX5-mediated DNA repair pathways in the emergence of cisplatin resistance in hypopharyngeal squamous cell carcinoma

作　　者：游雅婷 刘义钊 李维 余涛 郭睿 YOU Yating;LIU Yizhao;LI Wei;YU Tao;GUO Rui(Department of Otorhinolaryngology Head&Neck Surgery,Changde Hospital,Xiangya School of Medicine,Central South University,Hunan Changde 415000,China;Department of Otorhinolaryngology Head&Neck Surgery,Second Xiangya Hospital,Central South University,HunanChangsha 410028,China)

机构地区：[1]中南大学湘雅医学院附属常德医院耳鼻咽喉头颈外科,湖南常德415000 [2]中南大学湘雅二医院耳鼻咽喉头颈外科,湖南长沙410028

出　　处：《中国医院药学杂志》2024年第20期2376-2385,共10页Chinese Journal of Hospital Pharmacy

基　　金：湖南省卫生健康委科研计划项目(编号:202207012807)。

摘　　要：目的:阐明SOX5在下咽癌(hypopharyngeal carcinoma,HPC)中对顺铂耐药的影响及其作用机制。方法:生物信息学分析SOX5在HPC组织中的表达及富集通路,预测到SOX5的上游转录因子FOXM1,分析FOXM1在HPC组织中的表达,双荧光素酶和染色质免疫沉淀(ChIP)实验分析FOXM1对SOX5调控作用。qPCR检测SOX5和FOXM1在HPC细胞中的转录表达。CCK-8检测细胞活性、5-乙炔基-20脱氧尿苷检测细胞增殖能力、流式细胞术检测细胞凋亡率、彗星实验检测DNA损伤、免疫荧光检测γ-H2AX的表达、NHEJ报告分析检测DNA修复效率、Western blot检测DNA损伤修复相关蛋白表达、免疫组化检测相关蛋白的表达,并构建动物模型探究FOXM1表达对HPC肿瘤生长的影响。结果:SOX5在HPC中高表达,过表达SOX5介导DNA损伤修复从而促进HPC顺铂耐药性。FOXM1是SOX5的上游转录因子,且在HPC中高表达,回复实验发现敲低FOXM1表达能够拯救SOX5过表达通过DNA损伤修复对HPC顺铂耐药性的促进作用。体内实验结果表明敲低FOXM1表达联合顺铂治疗可显著降低异种移植肿瘤生长。结论:该研究表明FOXM1/SOX5轴介导的DNA损伤修复促进HPC顺铂耐药性,为改进化疗耐药后的治疗策略提供理论依据。OBJECTIVE To explore the effect of SOX5 on cisplatin(DDP)resistance in hypopharyngeal carcinoma(HPC)and elucidate its mechanism.METHODS The expression and enrichment pathway of SOX5 in HPC tissues were analyzed.The upstream transcription factor FOXM1 of SOX5 was predicted,the expression of FOXM1 in HPC tissues examined and binding relationship between the two was analyzed by dual luciferase and ChIP assays.The expression levels of SOX5 and FOXM1 in HPC cells were detected by quantitative polymerase chain reaction(qPCR).CCK-8 was utilized for detecting cell viability.Cell proliferation was detected by 5-ethynyl-2-deoxyuridine(EdU),apoptosis by flow cytometry,DNA damage by comet assay and expression ofγ-H2AX by immunofluorescence.Non-homologous DNA end joining(NHEJ)report analysis was utilized for detecting DNA repair efficiency and WB for detecting the expression of DNA damage repair-related proteins.The expression of related genes was detected by immunohistochemistry.An animal model was constructed for exploring the effect of FOXM1 expression on HPC tumor growth.RESULTS SOX5 was highly expressed in HPC.Overexpression of SOX5 mediated DNA damage repair and promoted DDP resistance in HPC.As an upstream transcription factor of SOX5,FOXM1 was highly expressed in HPC.Restoration experiments indicated that a knockdown of FOXM1 could attenuate the effect of SOX5 overexpres-sion on DDP resistance of HPC through DNA damage repair.In vivo assays showed that a knockdown of FOXM1 expression plus DDP dosing could significantly retard the growth of xenograft tumors.CONCLUSION FOXM1/SOX5 axis mediates DNA dam-age repair to promote HPC DDP resistance.The results of this study may help to optimize therapeutic strategies after chemo-therapy resistance.

关 键 词：FOXM1 SOX5 DNA损伤修复 下咽癌 顺铂耐药 

分 类 号：R965.2[医药卫生—药理学]