靶头修饰对PEG-PCL胶束在人宫颈癌细胞内转运行为的影响  

Effects of targeting modification on intracellular transportation of PEG-PCL micelles in human cervical cancer cells

作　　者：杨金津 于清华 于凌波 张亚东 梁冬芹 孙钰钰 王慧云[2] 崔亚男[2] YANG Jinjin;YU Qinghua;YU Lingbo;ZHANG Yadong;LIANG Dongqin;SUN Yuyu;WANG Huiyun;CUI Yanan(School of Pharmacy,Shandong Second Medical University,Shandong Weifang 261053,China;School of Pharmacy,Jining Medical University,Shandong Rizhao 276826,China;School of Pharmacy,Shandong First Medical University,Shandong Tai’an 271016,China)

机构地区：[1]山东第二医科大学药学院,山东潍坊261053 [2]济宁医学院药学院,山东日照276826 [3]山东第一医科大学药学院,山东泰安271016

出　　处：《中国药房》2024年第12期1431-1436,共6页China Pharmacy

基　　金：国家自然科学基金项目(No.81903553);山东省自然科学基金项目(No.ZR2017QH006);济宁医学院教育教学研究项目(No.yb202223)。

摘　　要：目的 研究转铁蛋白靶向肽T7(7pep)对聚乙二醇-聚己内酯(PEG-PCL)胶束在人宫颈癌HeLa细胞内转运行为的影响。方法 以香豆素-6(C6)为荧光指示探针,通过薄膜分散法制备包载有C6的PEG-PCL(PEG-PCL-C6)胶束以及靶头7pep修饰的PEG-PCL(7pep-PEG-PCL-C6)胶束。比较两种胶束的粒径、多分散指数及外观形态;比较两种胶束被HeLa细胞实时摄取的情况及其入胞后与早期内吞体(EE)、内吞循环室(ERC)、晚期内吞体(LE)的共定位情况。结果 PEG-PCL-C6和7pep-PEG-PCL-C6胶束的平均粒径分别为(75.0±2.3)、(82.0±1.5)nm,多分散指数分别为0.17±0.20、0.17±0.32,外观均为规整的圆球形。7pep-PEGPCL-C6胶束的入胞速度和入胞量均明显快/多于PEG-PCL-C6胶束。7pep-PEG-PCL-C6胶束比PEG-PCL-C6胶束能够更快地进入EE,而入胞后PEG-PCL-C6胶束进入ERC的速率较7pep-PEG-PCL-C6胶束快,且PEG-PCL-C6胶束和7pep-PEG-PCL-C6胶束在LE均有逐渐累积的趋势,但7pep-PEG-PCL-C6胶束入胞60 min时与LE的皮尔森系数、信号重叠比率、共定位比率均显著低于入胞30 min时(P<0.05或P<0.01)。结论 靶头7pep修饰可提高PEG-PCL-C6胶束的入胞速率和入胞量,还可改变其胞内转运行为。OBJECTIVE To study the effects of transferrin-targeting peptide T7(7pep) on intracellular transportation of polyethylene glycol-polycaprolactone(PEG-PCL) micelles in human cervical cancer HeLa cells.METHODS Using coumarin-6(C6) as fluorescent indicator probe,both coumarin-6(C6)-loaded PEG-PCL(PEG-PCL-C6) micelles and 7pep-modified PEGPCL(7pep-PEG-PCL-C6) micelles were prepared by film-dispersion method.The particle size,polydispersity index and appearance morphology were compared between two types of micelles;the real-time uptake of two types of micelles by HeLa cells was compared,and the colocalization of two types of micelles with early endosomes(EE),endocytic recycling compartments(ERC) and late endosomes(LE) after entry into the cells was observed.RESULTS The particle sizes of PEG-PCL-C6 and 7pep-PEG-PCLC6 micelles were(75.0±2.3)and(82.0±1.5)nm;the polymer dispersity indexes were 0.17±0.20 and 0.17±0.32,respectively,with a regular spherical appearance.The colocalization results showed that entry speed and amount of 7pep-PEG-PCL-C6 micelles were significantly faster/more than those of PEG-PCL-C6 micelles.7pep-PEG-PCL-C6 micelles entered EE faster than PEG-PCL-C6micelles,while PEG-PCL-C6 micelles entered ERC at a faster rate than 7pep-PEG-PCL-C6 micelles,and both PEG-PCL-C6micelles and 7pep-PEG-PCL-C6 micelles tended to accumulate gradually in LE;Pearson coefficient,signal overlap ratio,and colocalization ratio of 7pep-PEG-PCL-C6 micelles with LE were significantly lower 60 minutes after entering the cell than those 30 minutes after entering the cell(P<0.05 or P<0.01).CONCLUSIONS Targeting 7pep modification can increase the entry speed and amount of PEG-PCL-C6 micelles,and also alter their intracellular transportation behavior.

关 键 词：转铁蛋白靶向肽T7 胶束 转铁蛋白受体 靶向递送系统 细胞转运 

分 类 号：R943[医药卫生—药剂学]