基质金属蛋白酶-2在脑胶质瘤细胞中的表达及其分子探针体外磁共振成像的检测

摘要

Objective Observation of matrix metalloproteinase -2(MMP2)expression in the glioma cells,and study its molecular probes magnetic resonance imaging ability in vitro.Method The expression level of MMP2 was de-tected by Western blotting in various brain glioma cell line;USPIO-PEG-MMP2Ab was synthetized by conjugating USPIO-PEG with MMP2Ab.MTT colorimetric assay was used to assess the cytotoxicity of USPIO-PEG-MMP2Ab.The specific binding ability of USPIO-PEG and USPIO-PEG-MMP2Ab with MMP2 highly expressed glioma cells was detected by Prussian blue staining and MR imaging in vitro.Result All the tumor cells were positive for MMP2,there were sig-nificant differences compared with astrocytes;The synthetic molecular probe USPIO-PEG-MMP2Ab has no cytotoxicity in the range of concentration,and has the ability to bind specifically to C6 glioma cells.Prussian blue staining showed that at the same time and concentration conditions,USPIO-PEG-MMP2Ab group showed much more intracellular blue parti-cles than USPIO-PEG group.Magnetic resonance imaging showed that the SI value of the group USPIO-PEG-MMP2Ab cells under the same concentration condition decreased significantly,and compared with the USPIO-PEG group cells,the difference was statistically significant.Conclusion MMP2 is highly expressed in glioma cells, and the USPIO-PEG-MMP2Ab molecular probe has high activity and safety.It is targeted against glioma cell lines and lays the foundation in vivo experiments.%目的 观察基质金属蛋白酶-2(MMP2)在脑胶质瘤细胞中的表达,并检测其分子探针体外磁共振成像能力.方法 利用免疫印迹法检测 MMP2在多种脑胶质瘤细胞系的表达水平;利用碳二亚胺法合成USPIO-PEG-MMP2Ab分子探针,进行噻唑蓝比色法检测其细胞毒性,普鲁士蓝染色及体外磁共振成像检测USPIO-PEG及USPIO-PEG-MMP2Ab与MMP2高表达的脑胶质瘤细胞的特异性结合能力.结果 所有胶质瘤细胞均高表达MMP2,与星形胶质细胞相比,差异有统计学意义;合成的分子探针USPIO-PEG-MMP2Ab应用浓度范围内无明显细胞毒性,具有同C6脑胶质瘤细胞特异性结合的能力,普鲁士蓝染色结果显示在相同浓度及时间的条件下,USPIO-PEG-MMP2Ab 组较 USPIO-PEG 组蓝染颗粒更多、更明显;磁共振成像结果显示USPIO-PEG-MMP2Ab组细胞在相同浓度条件下SI值下降水平明显较高,与USPIO-PEG组细胞相比,差异有统计学意义.结论 MMP2在胶质瘤细胞中高表达,且USPIO-PEG-MMP2Ab分子探针活性高,安全性好,对脑胶质瘤细胞系具有靶向性.