荧光标记靶向纳米微球对胃癌细胞示踪作用的研究  

Tracing study of nanometer microspheres labeled with fluorescent dye on gastric cancer cells

作　　者：丁杰[1] 冯敏[1] 支文贤[2] 管文贤[1] 

机构地区：[1]南京大学医学院附属鼓楼医院普通外科,210008 [2]苏州大学附属儿童医院普通外科

出　　处：《中华胃肠外科杂志》2015年第2期171-176,共6页Chinese Journal of Gastrointestinal Surgery

基　　金：国家自然科学基金(81372364)；江苏省消化系统疾病临床医学中心(BL2012001)；南京市医学科技发展基金

摘　　要：目的：探讨近红外荧光染料（NIR-797）标记的靶向纳米微球对胃癌细胞的示踪作用，以期指导胃癌手术中的精准切除。方法采用酰胺化法将NH2-PEG-NH2、明胶酶作用底物肽段（Pep）及PCL-COOH合成靶向聚合物片段NH2-PEG-Pep-PCL及不含Pep的非靶向NH2-PEG-PCL，然后采用自组装法制备成空包纳米微球，通过化学反应将NIR-797标记到PEG末端。检测纳米微球的分子量、粒径、多分散性和表面电位；采用荧光显微镜观察非靶向和靶向纳米微球与人胃癌SGC-7901细胞的结合程度；同时采用多光谱成像系统检测静脉注射到荷瘤裸鼠体内不同时间点下两者在肿瘤部位的富集程度。结果靶向NH2-PEG-Pep-PCL及非靶向NH2-PEG-PCL的分子量和设计分子量相近，靶向纳米微球标记NIR-797前后的平均粒径分别为（124.1±2.1） nm和（129.7±2.8） nm，非靶向微球标记NIR-797前后的平均粒径分别为（120.6±2.6） nm和（124.3±2.9） nm，差异均无统计学意义（均P〉0.05）。靶向纳米微球在SGC-7901肿瘤细胞中和荷瘤裸鼠肿瘤部位的荧光强度均明显高于非靶向纳米微球。结论荧光标记靶向纳米微球对胃癌细胞有明显的靶向亲和力，并在肿瘤部位富集，可以很好地对肿瘤进行示踪，有望在临床上指导肿瘤的精准切除。Objective To explore the tracing role of nanometer microspheres tagged by fluorescent dye NIR-797 in gastric cancer cells in order to guide precise resection for gastric cancer. Methods Targeted polymer segment NH2-PEG-Pep-PCL and non-targeted polymer segment NH2-PEG-PCL（without Pep） were synthesized with NH2-PEG-NH2, gelatinase substrate peptides （Pep） and PCL-COOH by amidation method. Then nanometer microspheres were prepared by self-assemebly method. NIR-797 was linked into the end of PEG through chemical reaction. Molecular weight, particle size, polydispersity and surface potentials of the nanometer microspheres were detected. Uptake degree of human gastric SGC-7901 cells with targeted and non-targeted nanometer microspheres was observed under fluorescence microscope. Fluorescence intensity of above two nanometer microspheres in tumor sites of tumor-burdened nude mice after intravenous injection of nanometer microspheres at different time points was detected by multispectral imaging system. Results The molecular weight of NH2-PEG-Pep-PCL and NH2-PEG-PCL were similar to original design. The diameter of targeted nanometer microspheres before and after NIR-797-tagging was （124.1±2.1） nm and （129.7±2.8） nm, and the diameter of non-targeted ones before and after NIR-797-tagging was （120.6±2.6） nm and （124.3±2.9） nm （all P〉0.05）. Fluorescence intensity of SGC-7901 cells and tumor sites was significantly higher in the targeted＆amp;nbsp;nanometer microspheres group as compared to the non-targeted group. Conclusion Targeted nanometer microspheres tagged by flourescent dye have higher targeted affinity to gastric cancer cells and better enrichment in tumor site, indicating a good tracing of tumor and a guidance for accurate resection of tumor.

关 键 词：纳米微球 近红外荧光 胃肿瘤 示踪 裸鼠 

分 类 号：R735.2[医药卫生—肿瘤]