超声微泡包裹单纯疱疹病毒Ⅰ型胸苷激酶自杀基因靶向治疗小鼠肝癌的效果  

Effect of ultrasound microbubble carring herpes simplex virus thymidine kinase on hepatocellular carcinoma in mice

作　　者：周世骥[1] 刘长安[1] 龚建平[1] 刘作金[1] 唐勇[1] 李生伟[1] 徐越[1] 艾志国[1] 

机构地区：[1]重庆医科大学附属第二医院肝肌外科,400010

出　　处：《中华肝脏病杂志》2010年第4期276-279,共4页Chinese Journal of Hepatology

基　　金：国家自然科学基金（30872977）;重庆市卫生局科研项目（2008-2-192）;重庆市自然科学基金（2007BB0266）

摘　　要：目的观察超声微泡携单纯疱疹病毒Ⅰ型胸苷激酶（HSV1-TK）基因在小鼠肝癌细胞H22移植瘤组织中的转染效果及联合更昔洛韦（CCV）后对肿瘤的抑瘤效应。方法建立小鼠肝癌（H22）皮下移植瘤模型40只，随机分成4组，分别经鼠尾静脉注射入等量磷酸盐缓冲液（A组）、HSV1-TK（B组）、HSV1-TK（C组）、HSV1-TK＋做泡（D组），每次注射200μl，每隔3d注射1次，共注射3次；对C组、D组小鼠分别给予1MHz、2W／cm^2、5min超声辐照。首次注射HSV1 TK48h后采用Western blot检测肿瘤组织中TK蛋白的表达情况，且各治疗组分别从腹腔注射GCV0．2ml（100mg·kg^-1·d^-1），连续注射14d。测肿瘤大小，计算抑瘤率，治疗结束后处死动物行病理学检查。数据采用重复测量资料的方差分析，两两比较采用LSD—t法。结果在超声引导下微泡包裹的HSV1-TK基因可以准确定位于肿瘤组织并靶向释放，在处理组中均有TK蛋白的表达，其中D组表达量明显高于其他各组（P〈0．05）。抑瘤率在A组、B组、C组、D组中分别为0、3．90％±1．80％、22．70％±2．86％、41．25％±3．20％（C组、D组与B组相比，P值均〈0．05；D组与C组相比，P〈0．05）。HE染色结果显示D组较A组的坏死病变程度明娃减轻。结论超声辐照可破坏包裹HSV1-TK基因的微泡以定位释放，既增加了肝癌基因治疗的靶向性，又提高了外源基因的转染效率，从而增强了自杀基因对小鼠肝癌的杀伤效果。Objective To observe the effect of ultrasound microbubble carting herpes simplex virus thymidine kinase hepaocellular carcinoma in mice. Methods Kunming mice were inoculated subcutaneously with H22 tumor cells. 40 male mice bearing subcutaneous hepatoma were randomized into 4 groups： PBS （groupA）, HSV1-TK （group B）, HSV1-TK（group C）, and microbubble carring HSV1-TK （ group D） were injected into the tail vein every 3 days. Mice in group C and D were exposed to ultrasound. The expression of TK protein was detected by western blot. Ganciclovi （GCV） was intraperitoneally injected at a dose of 100 mg· kg· d^-1 in group B, group C and group D. The tumor size was measured every 2 days. Results TK gene could be injected precisely into hepatocellular carcinoma with ultrasound monitor, and the expression of TK protein was found in all 4 groups. Expression in group D was higher than others （P 〈 0.05）. The rate of tumor growth inhibition were 0 in group A, 3.90% ± 1.80% in group B, 22.70% ± 2.86% in group C, 41.25%± 3.20% in group D （group B vs group C, P 〈 0.05; group D vs group C, P 〈 0.05; groupD vs group B, P 〈 0.05）. Conclusions Ultrasound microbubble not only improve target gene therapy, but also enhance transfection efficiency.

关 键 词：癌 肝细胞 基因 HSV1-TK 超声微泡 

分 类 号：R735[医药卫生—肿瘤]