YTHDF3靶向PD-L1对肝细胞癌细胞增殖、迁移、侵袭和肿瘤干性的影响  

Effect of YTHDF3 targeted PD-L1 on proliferation,migration,invasion and tumor stemness in hepatocellular carcinoma

作　　者：宋雯茜 安亚军 朱婕 李远迪 苏敏 赵友波[1,2] 胡蓉 SONG Wenqian;AN Yajun;ZHU Jie;LI Yuandi;SU Min;ZHAO Youbo;HU Rong(Department of Histology and Embryology,School of Basic Medicine;National Joint Local Engineering Laboratory for Cell Engineering and Biomedicine Technique,Guizhou Province Key Laboratory of Regenerative Medicine,Key Laboratory of Adult Stem Cell Translational Research(Chinese Academy of Medical Sciences);Translational Medicine Research Center,Key Laboratory of Autoimmune Disease Research in Guizhou Colleges and Universities,Guizhou Medical University,Guizhou Guiyang 550025,China)

机构地区：[1]贵州医科大学基础医学院组织学与胚胎学教研室 [2]贵州医科大学细胞工程生物医药技术国家地方联合工程实验室,贵州省再生医学重点实验室,中国医学科学院成体干细胞转化研究重点实验室 [3]贵州医科大学转化医学研究中心,贵州省高等学校自身免疫病研究重点实验室,贵州贵阳550025

出　　处：《现代肿瘤医学》2024年第17期3164-3171,共8页Journal of Modern Oncology

基　　金：国家自然科学基金(编号:32260165);贵州省科技计划项目(编号:黔科合基础-ZK[2022]一般404);贵州省普通高等学校青年科技人才成长项目(编号:黔教合KY字[2022]244号)。

摘　　要：目的:探究YTHDF3在肝癌中的表达水平和YTHDF3对肝癌细胞增殖、迁移、侵袭和肿瘤干性的影响及可能涉及的靶点。方法:RT-PCR和Western blot检测临床肝癌组织和癌旁组织中YTHDF3的表达水平,使用正常肝细胞和肝癌细胞检测YTHDF3在mRNA和蛋白水平的表达。干扰YTHDF3在两组肝癌细胞中的表达,通过CCK-8、划痕实验、Transwell实验检测肝癌细胞活力、迁移和侵袭能力。干扰YTHDF3后观察肝癌细胞的成球能力和流式细胞术测定CD133干性标记的变化。最后,下调YTHDF3检测PD-L1在mRNA和蛋白水平的变化。结果:与癌旁组织相比,YTHDF3在肝癌组织中表达显著上调。与LO2细胞相比,两组肝癌细胞中YTHDF3的表达升高。干扰YTHDF3在肝癌细胞中的表达后,肝癌细胞的增殖、迁移、侵袭和成球能力减弱,CD133干性标记阳性率降低。下调YTHDF3在肝癌细胞中的表达,PD-L1的表达随之减少。结论:YTHDF3在肝癌中的高表达能够促进肝癌细胞的增殖、迁移、侵袭和肿瘤干性等特征,可能是作用于PD-L1来实现肝癌细胞免疫逃逸。Objective:To investigate the expression level of YTHDF3 in hepatocellular carcinoma and the effects of YTHDF3 on proliferation,migration,invasion and tumor stemness of hepatocellular carcinoma and possible targets involved.Methods:RT-PCR and Western blot were used to detect the expression level of YTHDF3 in clinical hepatocellular carcinoma tissues and paracancerous tissues,and the expression of YTHDF3 at the mRNA and protein levels was detected using normal hepatocytes and hepatocellular carcinoma cells.Interfering with the expression of YTHDF3 in hepatocellular carcinoma cells in both groups,hepatocellular carcinoma cell viability,migration and invasion ability were detected by CCK-8,scratch assay and Transwell assay.The sphere-forming ability of hepatocellular carcinoma cells and changes in CD133 stemness labeling determined by flow cytometry were observed after interfering with YTHDF3.Finally down-regulation of YTHDF3 detected changes in PD-L1 at mRNA and protein levels.Re sults:YTHDF3 expression was significantly up-regulated in hepatocellular carcinoma tissues compared with paracancerous tissues.Compared with LO2,YTHDF3 expression was elevated in hepatocellular carcinoma cells in both groups.Interference with YTHDF3 expression in hepatocellular carcinoma cells resulted in diminished proliferation,migration,invasion,and spheroid formation,and decreased CD1 3 3 stemness marker positivity.Down-regulation of YTHDF3 expression in hepatocellular carcinoma cells was followed by a decrease in PD-L1 expression.Conclusion:High expression of YTHDF3 in hepatocellular carcinoma promotes the proliferation,migration,invasion and stemness characteristics of hepatocellular carcinoma cells,and its role may be to achieve hepatocellular carcinoma cell immune escape by targeting PD-L1.

关 键 词：肝癌 YTHDF3 m^(6)A 肿瘤干性 免疫逃逸 

分 类 号：R735.7[医药卫生—肿瘤]