微泡造影剂联合超声介导KDR启动子-胸苷激酶基因靶向血管治疗小鼠肝癌  

Herpes simplex virus thymidine kinase gene therapy under the driving of KDR promoter for targeting angiogenesis in hepatocellular carcinoma of mice mediated by microbubble combined with ultrasound exposure

作　　者：聂芳[1] 徐辉雄[1] 吕明德[2] 王竹[1] 汤庆[3] 王瑛[1] 

机构地区：[1]中山大学附属第一医院超声科,广州510080 [2]中山大学附属第一医院肝胆外科 [3]广州医学院附属第一医院超声科

出　　处：《中华超声影像学杂志》2007年第5期448-452,共5页Chinese Journal of Ultrasonography

基　　金：国家自然科学基金（3047046730300082）;广东省科委重点攻关项目（2002C31108）;广东省自然科学基金（04009360）

摘　　要：目的 评估微泡造影剂SonoVue联合超声辐照介导KDR启动子-单纯疱疹病毒胸苷激酶基因（HSV—TK）系统（KDR—TK）结合更昔洛韦（GCV）靶向血管治疗小鼠肝癌的有效性。方法 建立小鼠（C57BL／6J）肝癌皮下移植瘤模型40只，随机分为4组，分别经尾静脉团注磷酸缓冲盐溶液（PBS）、KDR—TK、KDR—TK、KDR—TK＋SonoVue，后两组分别给予1MHz、2W／cm^2、5min的超声辐照，24h后治疗组分别腹腔注射GCV0．1ml（100mg·kg^-1·d^-1），连续10d，检测肿瘤大小，计算抑瘤率，28d后处死小鼠行肿瘤微血管密度及组织病理学检查。结果 抑瘤率在PBS组（A组）、KDR—TK／GCV组（B组）、KDR—TK＋超声／GCV组（C组）、KDR—TK＋超声＋SonoVue／GCV组（D组）分别为0%、3．6％±2．7％、21．4％±2．9V0和40．7％±4．5％（D组与B组，P〈0．01；D组与C组，P〈0．05；C组与B组，P〈0．05）；肿瘤微血管密度分别为48．8±5．1、44．2±6．4、27．4±3．2和12．3±1．4（B组与A组，P〉0．05；C组与A组，P〈0．05；D组与A组，P〈0．01；D组与C组，P〈0．05）；HE染色C组、D组坏死病变明显。结论 微泡造影剂增强超声辐照介导的KDR—TK基因靶向破坏小鼠肝癌微血管的效应，为肝癌的基因治疗提供一种可行、有效的方法。Objective To evaluate the efficacy of herpes simplex virus thymidine kinase（HSV-TK） under the control of kinase domain-containing receptor （KDR） promoter （KDR-TK） for targeting angiogenesis in hepatocellular carcinoma of mice mediated by microbubble combined with ultrasound. Methods C57BL/6J mice were inoculated subcutaneously in the middle of the right flank with hepa1-6 tumor cells line. A total of 40 female mice bearing subcutaneous hepatoma （≥5 mm in diameter） were randomized into 4 groups as follows： group A, PBS alone group B, KDR-TK group C, KDR-TK and ultrasound and group D,KDR-TK,ultrasound and SonoVue. There were 10 mice per group. The 150 /μl mixture of 60 μg plasmid KDR-TK and microbubbles/PBS was injected into the tail vein of each mouse. Pulse Doppler ultrasound was applied at a 1 MHz frequency with a 50% duty cycle and a 2.0 W/cm^2 output intensity for 5 min. Ganciclovi（GCV） was administered subsequently in each mouse daily by intraperitoneal injection at a dose of 0.1 ml（100 mg· kg^-1· d^-1 ） in group B,group C and group D. The tumor size was measured every 7 d till 28 d, then, all mice were killed to evaluated the microvessel density and the histopathological changes of tumors. Results The rate of tumor growth inhibition were 0% in group A, 3.6% ± 2.7% in group B,21.4% ± 2.9% in group C and 40.7% ± 4.5% in group D（group D vs group B, P 〈0. 01 ;group D vs group C, P 〈0. 05 group C vs group B, P 〈0. 05）. Microvessel density of tumor was 48.8 ± 5.1 in group A,44.2 ± 6.4 in group B,27.4 ± 3.2 in group C and 12.3 ±1.4 in group D respectively （group B vs group A.P〉0.05;group C vs group A.P〈0.05;group D vs group A,P〈0.01;group D vs group C,P〈0.05）There was more obvious necrosis of tumor tissue in group C and group D compared with that in group A and group B .Conclusions Microbubble contrast agent enhanced the effect of ultrasound mediated KDR-TK gene therapy on targeting angiogenesis in hepatoma of mice,which may provide a feasible and eff

关 键 词：超声检查 造影剂 肝肿瘤 基因疗法 

分 类 号：R686[医药卫生—骨科学]