叶酸靶向的纳米金载体构建及其对HeLa细胞杀伤作用的研究  

CONSTRUCTION OF FOLATE-TARGETED GOLD NANOPARTICLE VECTOR AND ITS KILLING EFFECT ON HELA CELLS

作　　者：刘树亮 葛南 崔莲花 LIU Shuliang , GE Nan, CUI Lianhua(School of Public Health, Qingdao University Medical College, Qingdao 266021, Chin)

机构地区：[1]青岛大学医学部公共卫生学院,山东青岛266021

出　　处：《精准医学杂志》2018年第3期274-278,282,共6页Journal of Precision Medicine

基　　金：山东省自然科学基金资助项目(ZR2011HM031)

摘　　要：目的构建叶酸靶向的纳米金载体并负载阿霉素(Au-BSA-DOX-FA),探讨其对人宫颈癌He La细胞的杀伤作用。方法以牛血清白蛋白为模板合成纳米金(Au-BSA),MTT法检测其细胞毒性。以在酸性环境下易断裂的顺乌头酸酐作为连接键结合DOX,达到p H敏感的可控释放药物效果。同时结合叶酸(FA)作为靶向分子,使药物可以靶向聚集于FA受体(FR)高表达的He La细胞表面。用表面FR低表达的人乳腺癌MCF-7细胞和人皮肤成纤维HSF细胞作为对照,比较Au-BSA-DOX-FA和DOX对两种癌细胞和正常细胞的杀伤作用。结果所合成的Au-BSA粒径约50 nm,稳定性好,对He La细胞、MCF-7细胞和HSF细胞无毒性作用。紫外吸收光谱表明,Au-BSADOX-FA已成功合成,并且在酸性环境中(p H为3.5、5.0)可以在12 h内持续释放DOX,释放率达到90%,在p H为6.5时DOX释放率约60%,而在p H为7.4、9.0时,释放率仅约20%。在对He La细胞的干预中,Au-BSA-DOX-FA的杀伤作用强于DOX的杀伤作用,差异有统计学意义(t=26.0,P<0.05);在对MCF-7细胞的干预中,Au-BSA-DOX-FA的杀伤作用与DOX的杀伤作用无明显差异;在对HSF细胞的干预中,Au-BSA-DOX-FA的杀伤作用弱于DOX的杀伤作用,差异有统计学意义(t=26.5,P<0.05)。结论 Au-BSA-DOX-FA作为FA靶向的载药平台对表面FR高表达的He La细胞有较好的杀伤作用。Objective To construct a folate（ FA）-targeted gold nanoparticle vector loaded with doxorubicin（ Au-BSA-DOXFA）,and to investigate its killing effect on human cervical cancer He La cells. Methods Gold nanoparticles were synthesized using bovine serum albumin as a template and MTT assay was used to evaluate cytotoxicity. These gold nanoparticles were conjugated with doxorubicin using cis-aconitic anhydride as a linker which was easily broken in an acidic environment to achieve the p H-sensitive controlled release of the drug. Meanwhile,FA was conjugated as a target molecule so that the drug can be targeted to the surface of He La cells with high expression of FA receptor. Human breast cancer MCF-7 cells and human skin fibroblast（ HSF） cells with low expression of FA receptor were selected as controls to investigate the killing effect of Au-BSA-DOX-FA versus DOX on these two cancer cell lines and normal cells. Results The synthesized Au-BSA gold nanoparticles had a diameter of 50 nm with good stability and had no cytotoxicity on He La cells,MCF-7 cells,and HSF cells. The UV absorption spectrum showed that Au-BSA-DOX-FA was successfully synthesized and it could release DOX continuously within 12 h in an acidic environment（ pH = 3.5 or 5.0）,with a release rate of 90%; the release rate was 60% at a pH of 6.5 and only about20% at a pH of 7.4 or 9.0. Au-BSA-DOX-FA had a significantly stronger killing effect on He La cells than DOX（ t = 26.0,P〈0.05）,while in MCF-7 cells,there was no significant difference in killing effect between Au-BSA-DOX-FA and DOX; in HSF cells,Au-BSA-DOX-FA had a significantly weaker killing effect than DOX（ t = 26.5,P〈0.05）. Conclusion As an FA-targeted drug-loaded platform,Au-BSADOX-FA has a good killing effect on He La cells with high expression of FA receptor on the surface.

关 键 词：纳米金 阿霉素 叶酸受体 HELA细胞 

分 类 号：R737.33[医药卫生—肿瘤]