Bian Ai-Na, Gao Yun-Hua, Tan Kai-Bin, Liu Ping, Zeng Gong-Jun, Zhang Xin, Liu Zheng
Department of Ultrasonic Diagnosis, Xinqiao Hospital, the Third Military Medical University, Chongqing 400037, China.
World J Gastroenterol. 2004 Dec 1;10(23):3424-7. doi: 10.3748/wjg.v10.i23.3424.
To prepare the human hepatocellular carcinoma-(HCC)-targeted liposome microbubbles and to investigate their immunological properties.
Human hepatocarcinoma specific monoclonal antibody HAb18 was attached to the surface of home-made liposome microbubbles by static attraction to prepare the targeted liposome microbubbles. The combination of HAb18 with liposome microbubbles was confirmed by the slide agglutination test and immunofluorescent assay. Their immunological activity was measured by ELISA. Rosette formation test, rosette formation blocking test and immun-ofluorescent assay were used to identify the specific binding of targeted liposome microbubbles to SMMC-7721 hepatoma cells, and cytotoxicity assay was used to detect their effect on human hepatocytes.
The targeted liposome microbubbles were positive in the slide agglutination test and immunofluorescent assay. ELISA indicated that the immunological activity of HAb18 on the liposome microbubbles was similar to that of free HAb18. SMMC-7721 cells were surrounded by the targeting liposome microbubbles to form rosettes, while the control SGC-7901 gastric cancer cells were not. Proliferation of SMMC-7721 cells and normal human hepatocytes was not influenced by the targeted liposome microbubbles.
The targeted liposome microbubbles with a high specific biological activity have been successfully prepared, which specifically bind to human hepatocarcinoma cells, and are non-cytotoxic to hepatocytes. These results indicate that the liposome microbubbles can be used as a HCC-targeted ultrasound contrast agent that may enhance ultrasound images and thus improve the diagnosis of HCC, especially at the early stage.
制备人肝细胞癌(HCC)靶向脂质体微泡并研究其免疫学特性。
通过静电吸附将人肝癌特异性单克隆抗体HAb18连接到自制脂质体微泡表面,制备靶向脂质体微泡。通过玻片凝集试验和免疫荧光测定法确认HAb18与脂质体微泡的结合。用ELISA测定其免疫活性。采用玫瑰花结形成试验、玫瑰花结形成阻断试验和免疫荧光测定法鉴定靶向脂质体微泡与SMMC - 7721肝癌细胞的特异性结合,并用细胞毒性试验检测其对人肝细胞的作用。
靶向脂质体微泡在玻片凝集试验和免疫荧光测定中呈阳性。ELISA表明脂质体微泡上HAb18的免疫活性与游离HAb18相似。SMMC - 7721细胞被靶向脂质体微泡包围形成玫瑰花结,而对照的SGC - 7901胃癌细胞则未形成。靶向脂质体微泡不影响SMMC - 7721细胞和正常人肝细胞的增殖。
已成功制备出具有高特异性生物活性的靶向脂质体微泡,其能特异性结合人肝癌细胞,对肝细胞无细胞毒性。这些结果表明脂质体微泡可作为一种HCC靶向超声造影剂,可能增强超声图像,从而改善HCC的诊断,尤其是早期诊断。