瑞戈非尼联合PD-L1单抗对小鼠肝癌移植瘤的作用及机制  

Effects and mechanisms of regorafenib combined with PD-L1 antibody on liver cancer xenograft in mice

作　　者：倪明立 潘威 徐倩 NI Mingli;PAN Wei;XU Qian(The Second Department of Oncology,Luoyang Central Hospital Affiliated to Zhengzhou University,Luoyang,Henan 471000,China)

机构地区：[1]郑州大学附属洛阳中心医院肿瘤二科,河南洛阳471000

出　　处：《中国普通外科杂志》2024年第7期1091-1099,共9页China Journal of General Surgery

基　　金：河南省科学技术厅科技发展计划基金资助项目(202102310048)。

摘　　要：背景与目的:瑞戈非尼治疗肝癌不可避免的存在毒副作用,而且治疗有效率有限。细胞程序性死亡-配体1(PD-L1)单抗可有效阻断“肿瘤免疫逃逸机制”,发挥显著抗肿瘤作用。因此,本研究探讨瑞戈非尼联合PD-L1单抗对肝癌移植瘤小鼠的抗癌作用。方法:将Balb/C裸小鼠建立肝癌移植瘤模型后,分别给予瑞戈非尼(瑞戈非尼组)、阿替利珠单抗(PD-L1单抗组)、瑞戈非尼^(+)阿替利珠单抗(联合组)、生理盐水(模型组)处理,连续4周,期间动态测量各组肿瘤体积。4周后,剥离各组小鼠移植瘤,分别用流式细胞术、TUNEL法、HE染色检测肿瘤组织CD4^(+)和CD8^(+)细胞浸润情况、细胞凋亡率及形态特征,并用qRT-PCR和Western blot检测肿瘤组织CD31、血管内皮生长因子(VEGF)、增殖细胞核抗原(Ki-67)、B淋巴细胞瘤-2基因(Bcl-2)、Bax表达。结果:给药前4组小鼠肿瘤体积差异无统计学意义(P>0.05),给药1、2、3、4周时,各给药组小鼠肿瘤体积均明显小于模型组,且联合组的肿瘤体积明显小于两个单药组(均P<0.05),但两个单药组间肿瘤体积无明显差异(均P>0.05)。各给药组肿瘤组织中CD4^(+)与CD8^(+)细胞比例均较模型组明显升高,且升高程度依次为瑞戈非尼组<PD-L1单抗组<联合组(均P<0.05)。各给药组肿瘤组织细胞凋亡率均较模型组明显升高,且升高程度依次为瑞戈非尼组<PD-L1单抗组<联合组(均P<0.05)。HE染色显示,模型组肿瘤细胞核大、深染、排列密集,各给药组肿瘤细胞均出现不同程度的核皱缩、细胞数量减少,片状坏死等改变,联合组改变最为明显。与模型组比较,各给药组肿瘤组织CD31、VEGF、Ki-67和Bcl-2 mRNA与蛋白表达水平明显降低,Bax mRNA与蛋白表达水平明显升高,且联合组的以上指标变化程度明显大于两个单药组,但两个单药组间差异均无统计学意义(均P>0.05)。结论:瑞戈非尼联合PD-L1单抗可有效抑制肝癌生长,且强�Background and Aims:Regorafenib treatment for liver cancer inevitably presents side effects and limited efficacy.Programmed death-ligand 1(PD-L1)monoclonal antibody can effectively block the"tumor immune escape mechanism"and exert significant anti-tumor effects.Therefore,this study was conducted to explore the anti-cancer effect of regorafenib combined with PD-L1 monoclonal antibody in a mouse model of transplanted liver cancer.Methods:After establishing a liver cancer transplant model in Balb/C nude mice,the mice were treated with regorafenib(regorafenib group),atezolizumab(PD-L1 antibody group),regorafenib^(+)atezolizumab(combination group),or saline(model group)for 4 weeks.Tumor volumes were dynamically measured in each group during the treatment period.After 4 weeks,the transplanted tumors were excised from the mice,and flow cytometry,TUNEL assay,and HE staining were used to detect CD4^(+)and CD8^(+)cell infiltration,apoptosis rate,and morphological characteristics of the tumor tissues.Additionally,qRT-PCR and Western blot were employed to detect the expression of CD31,vascular endothelial growth factor(VEGF),proliferating cell nuclear antigen(Ki-67),B-cell lymphoma 2(Bcl-2),and Bax in the tumor tissues.Results:There was no statistically significant difference in tumor volumes among the four groups of mice before treatment(P>0.05).At weeks 1,2,3,and 4 of treatment,the tumor volumes in each treated group were significantly smaller than those in the model group,and the tumor volume in the combination group was significantly smaller than those in the two monotherapy groups(all P<0.05).However,there was no significant difference in tumor volume between the two monotherapy groups(all P>0.05).The proportion of CD4^(+)and CD8^(+)cells in tumor tissues was significantly higher in each treated group than those in the model group,with the increase being in the order of regorafenib group<PD-L1 antibody group<combination group(all P<0.05).The apoptosis rate of tumor cells was significantly higher in each treated group co

关 键 词：肝肿瘤 肿瘤移植 小鼠 瑞戈非尼 阿替利珠单抗 

分 类 号：R735.7[医药卫生—肿瘤]