DNA-PK抑制剂联合溶瘤病毒增强对肝癌的抗肿瘤活性研究  

DNA-PK inhibitor combined with oncolytic virus enhances anti-tumor activity of liver cancer

作　　者：吴育民[1] 杜端明[1] 刘春霖[1] 洪跃飞 江磊昌 罗伟芝 彭尧 JOHN Bell WU Yu-Min;DU Duan-Ming;LIU Chun-Lin;HONG Yue-Fei;JIANG Lei-Chang;LUO Wei-Zhi;PENG Yao;JOHN Bell(Department of Interventional Therapy,Shenzhen Second People's Hospital,Shenzhen 518000,China)

机构地区：[1]深圳市第二人民医院介入治疗科,深圳518000 [2]广州医科大学基础医学院,广州510182 [3]渥太华健康研究所,渥太华999040

出　　处：《中国免疫学杂志》2021年第18期2231-2234,2239,共5页Chinese Journal of Immunology

基　　金：广东省自然科学基金项目(2018A030313370);吴阶平医学基金会临床科研专项资助基金(320.6750.19073-17)资助。

摘　　要：目的:寻找协同增强VSVΔ51溶瘤病毒(OV)对肝癌杀伤作用的小分子化合物并探索其协同机制。方法:通过将不同机制的抗肿瘤活性的小分子化合物与VSVΔ51联合,筛选对肝癌细胞裂解能力最强的药物组合;通过体内抑瘤能力实验检测联合疗法的体内抗肿瘤活性;通过电镜观测联合疗法处理后内质网的肿胀程度及Western blot检测肿瘤组织中凋亡相关蛋白的水平,探索联合疗法协同增效的机制。结果:细胞存活实验表明,DNA-PK抑制剂M3814与VSVΔ51联合后较M3814单独给药细胞毒性明显增强,沉默靶细胞DNA-PKcs基因后也能显著增强VSVΔ51的肿瘤裂解能力(P<0.0001);体内实验表明,M3814与VSVΔ51联合治疗较单独治疗而言肿瘤抑制能力显著增强(P<0.001);电镜观测结果显示M3814与VSVΔ51联合处理靶细胞后内质网肿胀程度增加(P<0.001),Western blot检测结果表明,M3814与VSVΔ51联合治疗后的肿瘤组织中凋亡相关蛋白表达增加(P<0.001)。结论:M3814联合VSVΔ51可以增强内质网应激诱导肝癌细胞凋亡,有可能为肝癌的治疗提供新的方案。Objective:To find the small molecular compounds that synergistically enhance the cytotoxicity effect of VSVΔ51 oncolytic virus(OV)on hepatocellular carcinoma and explore its synergistic mechanism.Methods:By combining the small molecular compounds with different mechanisms of anti-tumor activity with VSVΔ51,the combinatory therapy with the strongest lytic ability to hepatoma cells were screened,and the anti-tumor activity of the combinatory therapy in vivo was detected by tumor inhibition experiment in vivo.The swelling degree of the endoplasmic reticulum was observed by electron microscope and the level of apoptosis-related proteins in tumor tissue was detected by Western blot to explore the mechanism of the synergism of combined therapy.Results:Cell survival experiment showed that the cytotoxicity of DNA-PK inhibitor M3814 combined with VSVΔ51 was significantly stronger than that of M3814 alone,and the tumor lytic ability of VSVΔ51 was also significantly enhanced by silencing target cell DNA-PKcs gene(P<0.0001).The in vivo experiment showed that the tumor inhibitory ability of M3814 combined with VSVΔ51 was significantly stronger than that of M3814 alone(P<0.001),and the electron microscopic observation showed that the swelling of endoplasmic reticulum increased after the target cells were treated with M3814 and VSVΔ51(P<0.001).Western blot results showed that the expression of apoptosis-related proteins in tumor tissue was increased after treatment with M3814 and VSVΔ51(P<0.001).Conclusion:M3814 combined with VSVΔ51 can enhance the apoptosis of hepatoma cells induced by endoplasmic reticulum stress,which may provide a new strategy for the treatment of hepatocellular carcinoma.

关 键 词：肝细胞癌 溶瘤病毒 DNA-PK 肿瘤免疫治疗 

分 类 号：R735.7[医药卫生—肿瘤] R730.54[医药卫生—临床医学]