功能化石墨烯载hTERTsiRNA的制备及其对宫颈癌Hela细胞的影响  

Preparation of hTERTsiRNA Loaded Functionalized Graphene and Its Influence on Hela

作　　者：牛高丽[1] 朱建平 徐鸣 杨晓英[3] 温玉库[1] 陈永胜[4] 

机构地区：[1]辽宁医学院第一附属医院,辽宁沈阳121001 [2]无锡东方肿瘤医院生物治疗中心,江苏无锡214028 [3]天津医科大学天津市临床药物关键技术重点实验室,天津300070 [4]南开大学化学学院,纳米科学与技术中心功能聚合物材料重点实验室,天津300071

出　　处：《南开大学学报（自然科学版）》2013年第3期35-40,共6页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：中国博士后科学基金(201104289)

摘　　要：以氧化石墨烯(GrapheneOxide,GO)为基质,制备了氧化石墨烯-聚乙二醇-叶酸(GO-PEG-FA)纳米载体,借助于通过π-π相互作用负载在GO表面的胺甲基芘盐酸盐(PyNH2),介导人端粒酶反转录酶siRNA(hTERTsiRNA)转染入宫颈癌Hela细胞.GO-PEG-FA-PyNH2-hTERTsiRNA纳米基因复合物被成功制备,并通过红外光谱和紫外光谱进行表征.荧光显微镜观察该纳米载体的转染效率明显优于裸siRNA组,Western blot实验检测Hela细胞hTERT蛋白表达显著降低,MTT实验观察该载体对细胞无毒,其共载hTERTsiRNA和抗肿瘤药阿霉素后对Hela细胞生长的抑制具有协同作用.因而该功能化氧化石墨烯可以作为靶向基因载体,有效传递siRNA及化疗药物进入肿瘤细胞发挥作用.Graphene oxide - poly（ethylene glycol） - folio acid （GO-PEG-FA） was prepared with GO using as the matrix. Human telomerase reverse transcriptase small interfering RNA （hTERTsiRNA）was transfected into Hela cells by the functionalized GO with the aid of 1- pyrenemethylamine hydrochloride（PyNH2）loaded on the surface of GO via rt-n stacking. GO- PEG-FA-PyNHz-hTERTsiRNA nanocomplex was prepared successfully and characterized by a Fourier transform infrared spectroscopy and ultra-visible absorption spectrum. The transfec- tion efficiency of siRNA on these nanocarriers was obvious better than naked siRNA by the ob- servation of the confocal fluorescence microscope. The GO-PEG-FA-PyNHz transfected hTERTsiRNA clearly downregulated the protein expression level by western blot analysis. These nanocarriers were nontoxicity toward cells by MTT assays. After hTERTsiRNA and anticancer drug doxorubicin hydrochloride was co-delivered into Hela cells by GO-PEG-FA- PyNHz, they exhibit a synergistic effect to inhibit the growth of Hela. Therefore, the function- alized GO can be used as targeted gene carriers and delivered siRNA and anticancer drugs into tumor cells effectively.

关 键 词：石墨烯 叶酸 SIRNA HELA 

分 类 号：Q78[生物学—分子生物学]