超声微泡携MAGL-shRNA 靶向释放对大鼠肝细胞肝癌转移作用的实验研究.docVIP

超声微泡携MAGL-shRNA靶向释放对大鼠肝细胞肝癌转移作用的实验研究 陈懿1 王晓波2 张俊勇1 连峥嵘1 龚建平1(1400010 重庆，重庆医科大学附属第二医院肝胆外科 ; 2 408000 重庆，重庆市涪陵中心医院肝胆外科) [摘要]目的 观察超声微泡携单酰基甘油脂肪酶沉默基因(monoacylglycerol lipase short hairpin RNA, MAGL-shRNA)在大鼠肝细胞肝癌(hepatocellular carcinoma, HCC)组织中转染及对HCC转移的作用。方法 建立大鼠肝细胞肝癌模型，病理解剖和二维超声验证肝脏成瘤情况。抽取48只SD大鼠随机分成四组，分别为PBS液组、MAGL-shRNA质粒微泡组(MAGL-shRNA+microbubble, MB)、空白质粒微泡+超声辐照组(microbubbles+ultrasound, MB+US)、MAGL-shRNA质粒微泡+超声辐照组(MAGL-shRNA+MB+US)。每只注射1ml，对MB+US组和MAGL-shRNA+MB+US组大鼠肝区同时给予超声辐照，辐照条件为300kHz，2W/cm2，辐照10s，间隔10s，共20min。Western-Blot检测大鼠HCC组织MAGL蛋白的表达，免疫组化检测MAGL和基质金属蛋白酶2(matrixmetalloproteinase-2, MMP-2)的表达。比较各组动物的肿瘤转移情况。结果 MAGL在HCC组织中表达明显高于正常肝组织(P0.05); 微泡携MAGL-shRNA可以在HCC组织被超声辐照击破后靶向释放，在各组HCC组织中均有MAGL蛋白的表达，其中MAGL-shRNA+MB+US组表达量明显低于其它组(P0.01); 免疫组化检测MAGL-shRNA+MB+US组MMP-2表达均低于其它组(P0.01); 各组动物均见肿瘤转移，但MAGL-shRNA+MB+US组转移率最低(P0.01)。结论 超声辐照可破坏携MAGL-shRNA的微泡使之靶向释放并增强了MAGL-shRNA的转染效率，MAGL-MMP-2通路可能与HCC的转移相关。 [关键词] 肝细胞肝癌; 单酰基甘油脂肪酶; 基因; 超声微泡; 转移 Experimental research of ultrasound microbubble carrying MAGL-shRNA on hepatocellular carcinoma metastasis in rat CHEN Yi1 WANG Xiaobo2 ZHANG Junyong1 LIAN Zhengrong1 GONG Jianping1 (1Department of Hepatobiliary Surgery, Secondary Affiliated Hospital, Chongqing Medical University ，Chongqing, 400010, China; 2Department of Hepatobiliary Surgery, Central Hospital of Fuling district，Chongqing 408000, China) [Abstract] Objective To observe the transfection of monoacylglycerol lipase shRNA and the effect of inhibiting metastasis of hepatocellular carcinoma in rat by using ultrasound-targeted microbubble destruction. Methods Liver tumor models of rats were established. The successful establishment of tumor was testified by pathologic biopsy and two-dimensional ultrosound. 48 rats were randomly divided into 4 groups: phosphate-buffered solution group, lipid microbubbles loaded MAGL-shRNA (MAGL-shRNA+Microbubble group), pure lipid microbubbles+ultrasound (MB+US group), and lipid microbubbles loaded MAGL-shRNA+