超声介导载sPD-1和miR-206基因纳米微泡协同抑制小鼠H22肝癌皮下移植瘤  

Ultrasound-mediated sPD-1 and miR-206 gene nanoscale microbubbles synergistic inhibition of H22 hepatoma subcutaneous xenografts in mice

作　　者：谭妍迪 赵云[1] 刘朝奇[1] 周军[2] 郑智唯 马瑶[1] 姜矜君 TAN Yandi;ZHAO Yun;LIU Chaoqi;ZHOU Jun;ZHENG Zhiwei;MA Yao;JIANG Jinjun(Medical College of China Three Goreges University,Yichang 443002,China;Department of Ultrasonography,Yichang Central People's Hospital,Yichang 443000,China)

机构地区：[1]三峡大学医学院,湖北宜昌443002 [2]宜昌市中心人民医院超声科,湖北宜昌443000

出　　处：《中国医学影像技术》2019年第9期1315-1320,共6页Chinese Journal of Medical Imaging Technology

基　　金：宜昌市重点实验室(三峡大学)开放基金项目(2017KNX06);三峡大学学位论文培优基金项目(2019SSPY111)

摘　　要：目的探讨超声介导载可溶性程序性死亡因子1(sPD-1)联合miR-206纳米微泡对小鼠H22肝癌皮下移植瘤的干预效果。方法自制载sPD-1和miR-206基因的纳米微泡,构建H22肝细胞癌皮下移植瘤小鼠模型,并将模型小鼠随机分为模型组、空微泡组、miR-206微泡组、sPD-1微泡组、联合组(给予miR-206微泡组及sPD-1微泡),分别每2天经尾静脉给予生理盐水及相应的微泡,每次注射后给予超声辐照瘤体治疗1次。5次治疗后取小鼠肿瘤组织,测量肿瘤质量及体积,计算肿瘤体积及质量抑瘤率,HE染色观察肿瘤组织病理变化,免疫组织化学法检测小鼠肿瘤组织Bax、Bcl-2蛋白表达,半定量PCR检测Bax、Bcl-2、c-met、γ干扰素(IFN-γ)、程序性死亡因子-1配体(PD-L1)mRNA表达,实时荧光定量PCR检测miR-206表达。结果制备的纳米微泡呈球形,分布均一。与模型组比较,各组肿瘤体积、肿瘤质量均降低,体积抑瘤率及质量抑瘤率均升高(P均<0.05);上述变化以联合组为著(P均<0.05)。与模型组比较,其余各组Bax蛋白和mRNA均高表达、Bcl-2蛋白和mRNA均低表达,以联合组为著(P均<0.01)。各组小鼠肿瘤组织Bax、Bcl-2、c-met、PD-L1、IFN-γ、miR-206mRNA表达差异均有统计学意义(P均<0.01)。结论超声介导载sPD-1和miR-206纳米微泡可协同抑制小鼠H22肝癌皮下移植瘤生长。Objective To investigate the effect of ultrasound-mediated soluble programmed cell death 1 receptor (sPD-1) and miR-206 loaded nanoscale microbubbles on H22 hepatoma subcutaneous xenografts in mice.Methods sPD-1 and miR-206 loaded nanoscale microbubbles were prepared.The mice models of H22 hepatoma xenografts were established and randomly divided into model group,microbubble control group,miR-206 microbubble group,sPD-1 microbubble group and combined group (miR-206 and sPD-1 microbubbles).Mice in each group (each n =8) were treated with normal saline or corresponding nanoscale microbubbles every 2 days by injection via tail vein,and then irradiated by ultrasound once after every injection.Tumor tissues were obtained after being treated 5 times.Tumor volume and quality were measured,the volume and quality inhibitory rates were calculated.HE staining was used to observe pathological changes of the tumors.The expressions of Bcl-2,Bax proteins were detected by immunohistochemistry.The expressions of Bcl-2,Bax,c-met,interferon-γ(IFN-γ) and programmed cell death 1 receptor ligand (PD-L1) mRNA were detected with RT-PCR.Quantitative real-time fluorescence PCR was used to detect the expression of miR-206. Results The nanoscale bubbles were spherical and distributed uniformly.Compared with model group,tumor volume and quality decreased in other groups,and the volume and quality inhibitory rates increased (all P <0.05),especially in combined group (all P <0.05).Compared with model group,the Bax protein and mRNA expressions both increased,whereas the Bcl-2 protein and mRNA expressions decreased in other groups,especially in combined group (all P <0.01).There were significant differences of Bax,Bcl-2,c-met,PD-L1,IFN-γ and miR-206 mRNA in tumor tissues among each group (all P <0.01).Conclusion Ultrasound-mediated sPD-1 combine miR-206 loaded nanoscale microbubbles can synergistically inhibit H22 hepatoma xenografts in mice.

关 键 词：癌 肝细胞 微气泡 纳米粒子 微RNAS 超声检查 

分 类 号：R735.7[医药卫生—肿瘤] R445.1[医药卫生—临床医学]