PD-L1在小鼠肝内胆管癌模型中的实验研究

Experimental study of PD-L1 in mouse model of intrahepatic cholangiocarcinoma

目的通过水流动力学方法建立小鼠肝内胆管癌(ICC)模型;观察并分析PD-L1、IFN-γ、TNF-α、VEGF分子在ICC小鼠模型中的表达;通过小动物活体光学成像系统进行生物发光检测,验证PD-L1抗体对小鼠ICC模型的治疗效果。方法提取诱癌质粒,利用水流动力学方法注射至小鼠体内,建立小鼠ICC模型;通过小动物活体光学成像系统以及小动物B超观察肿瘤模型构建结果,取荷瘤小鼠ICC肿瘤组织进行HE染色,证明其造模情况;取正常肝脏组织及荷瘤小鼠ICC肿瘤组织,检测PD-L1及肿瘤微环境相关因子IFN-γ、TNF-α、VEGF浓度,运用Pearson相关系数法分析PD-L1与IFN-γ、TNF-α、VEGF之间的相关性;对ICC小鼠给予PD-L1抗体治疗,通过小动物活体光学成像系统观察PD-L1抗体治疗效果。结果4周后小鼠全部成瘤,成瘤率100%,经小动物活体光学成像系统观测,荷瘤小鼠肝脏部位显示强生物发光;经小动物B超观测,荷瘤小鼠肝脏部位可见液性暗区及肿块回声;HE染色结果显示,荷瘤小鼠肿瘤组织切片中肿瘤细胞呈胆管样排列。ELISA结果表明PD-L分子在ICC小鼠中呈高表达,其表达与肿瘤微环境相关因子IFN-γ、TNF-α、VEGF具有线性关系。经小动物活体光学成像系统检测,荷瘤小鼠经PD-L1抗体治疗后肝脏部位生物发光信号减弱。结论小鼠ICC模型构建成功,PD-L1分子在ICC小鼠中呈高表达,其表达与ICC肿瘤微环境细胞因子相关;PD-L1抗体对小鼠ICC模型具有一定治疗作用。

Objective Intrahepatic cholangiocarcinoma(ICC)mouse model was established by hydrodynamic method.The expression of PD-L1,IFN-γ,TNF-α,and VEGF numerator was observed analyzed.To validate the efficacy of PD-L1 antibody on ICC mouse model by using in vivo optical imaging system for small animals.Methods The tumor-inducing plasmid has been extracted,and was injected into mouse by using hydrodynamic method to establish ICC mouse model.The animal imaging system and animal B-ultrasound was used to observe the results of the tumor model construction,and the ICC tumor tissues of tumor-bearing mice were stained with HE to prove their modeling status.Normal liver tissues and ICC tumor tissues of tumor-bearing mice were taken to detect concentration of PD-L1 and tumor micro-environment related factors IFN-γ,TNF-α,VEGF.Pearson correlation coefficient method was to analyze the correlation between PD-L1 and IFN-γ,TNF-α,VEGF.ICC model mice were given PD-L1 antibody for treatment,in vivo optical imaging system for small animals was applied to observe the therapeutic effect of PD-L1 antibody.Results 4 weeks later,all mice had tumors,with a tumor formation rate of 100%.The liver of the tumor-bearing mice showed strong bioluminescence when observed by the small animal imaging system.The results of B-ultrasound indicated that liquid dark areas and tumor echo can be seen in the liver of tumor-bearing mice.HE staining results showed that the tumor cells in the tumor tissue section of the tumor-bearing mice were arranged like a bile duct.ELISA results showed that PD-L1 molecule was highly expressed in ICC mice,and its expression had a linear relationship with tumor micro-environment-related factors IFN-γ,TNF-αand VEGF.In the tumor-bearing mice that were treated with PD-L1 antibody by using animal imaging system,the bioluminescence signal in the liver was weakened.Conclusions The ICC mouse model was successfully constructed.PD-L1 molecule is highly expressed in the ICC mouse model,and its expression is related to ICC tumor micro-environment cytokines.PD-L1 antibody has a therapeutic effect on the ICC mouse model.