同步辐射成像技术应用于HCCLM3细胞裸鼠肝脏移植瘤新生血管形态学研究的初步实验  

Hepatocellular carcinoma and angiolgenesis imaging using synchrotron radiation

作　　者：李蓓蕾[1] 周波[2] 吴伟忠[3] 张一秋[1] 杜国浩[4] 石洪成[1] 陈绍亮[1] 

机构地区：[1]复旦大学附属中山医院核医学科,上海200032 [2]复旦大学附属中山医院核介入科,上海200032 [3]复旦大学附属中山医院肝癌研究所,上海200032 [4]中国科学院上海应用物理研究所

出　　处：《中华肝脏病杂志》2013年第9期684-687,共4页Chinese Journal of Hepatology

基　　金：国家重点基础研究发展计划（“973”计划,2010CB834305）

摘　　要：目的应用同步辐射微血管造影技术进行裸鼠离体肝细胞癌肿瘤新生血管的形态学研究。方法建立高转移人肝癌细胞HCCLM3裸鼠肝脏移植瘤模型。肝脏移植瘤模型建立第28天，取模型鼠6只，随机分为2组（每组3只）。第1组裸鼠经腹腔内注射戊巴比妥钠麻醉后开腹，经下腔静脉留置管，手动推注硫酸钡悬浮液，直至肝脏及肝肿瘤表面血管呈白色，结扎肝脏的血管和胆管后切除肝脏，浸泡在甲醛溶液里，准备成像。离体裸鼠肝癌新生血管成像在上海光源（SSRF）X射线成像与生物医学应用光束线站（BLl3W）进行。第2组裸鼠腹腔内注射戊巴比妥钠麻醉后，开腹取肝肿瘤标本，行病理学检查，包括HE染色及免疫组织化学CD31、CD34和F8染色。结果同步辐射微血管造影结合显微CT获得了高质量的肿瘤新生血管图像，图像显示正常肝血管结构被迅速生长的肿瘤组织取代，可以清晰观察到肿瘤新生血管的形态特征及生长状况，肿瘤周围血管丰富、扭曲，瘤内血管稀疏，与免疫组织化光学显微图像所示肿瘤新生血管分布特征相似。同步辐射成像技术能分辨肿瘤内部最细的血管直径约20μm。结论采用硫酸钡作为造影剂，利用同步辐射微血管造影技术显示裸鼠肝肿瘤新生血管的方法可行。Objective To investigate the potential utility of microangiography with synchrotron radiation to detect murine hepatocellular carcinoma （HCC） angiogenesis using an ex vivo model system. Methods An HCC xenograft model was established by implanting HCCLM3 cells into male mice livers （a = 6）. Twenty-eight days later, three of the mice were randomly selected for barium sulfate infusion into the liver and tumor via the inferior vena cava followed by ligation of the arteries, veins and common bile duct; the remaining three mice were left untreated and served as controls. All mice were sacrificed to collect livers for analysis using the BL13W beamline X-ray imager （Shanghai Synchrotron Radiation Facility, China）. In addition, the tumor vasculature was evaluated by immunostaining of formalin-fixed tissues for CD31, CD34, and F8. Results High resolution images of tumor angiogenesis were acquired and image analysis indicated that the normal blood vessels had been displaced by the fast growing tumors. Abundant and tortuous tumor angiogenesis in the tumor periphery area and sparse angiogenesisinside the tumor were also visualized clearly. These features were similar to the immunohistological results. The smallest tumor vessels visualized were approximately 20 μm in diameter. Conclusion Microangiography with synchrotron radiation using barium sulfate as contrast agent is a viable imaging strategy for tumor angiogenesis.

关 键 词：癌 肝细胞 新生血管化 病理性 血管造影术 体层摄影术 X线 同步辐射 

分 类 号：R735.7[医药卫生—肿瘤]