ge11肽介导的靶向脂质体抗肿瘤作用及摄取机制的分析 word格式.docxVIP

Study of the anti-tumor effect and cellular uptake mechanisms of GE11 peptide modified targeting liposomesAbstractObjective: In this study, we constructed peptide GE11 decorated DOX-loaded liposomes in order to actively target to EGFR over-expressed A549 cells. The in vitro cell experiments were used to evaluate the targeting efficiency, cellular cytotoxicity, cellular uptake. We also evalued the tumor accumulation of GE11 modified liposomes in A549 tumor-bearing nude mice. At last, the cellular uptake mechanism of GE11 decorated DOX-loaded liposomes was studied. All of these experiments aimed to provide evidences for this active targeting drug delivery system against NSCLC.Methods: The DSPE-PEG2000-GE11 were synthesized by addition reaction between DSPE-PEG2000-Mal and Cys-GE11, then the structure of the product was characterized by 1H-NMR and FTIR and the conjugation efficiency of GE11 to DSPE-PEG2000-Mal was quantitatively determined by HPLC. The GE11-LP/DOX was prepared by combination of the thin film hydration and post insertion method. Then the particle size, zeta potential, morphology and encapsulation efficiency of liposomes were evaluated by DLS, TEM and HPLC. The MTT assay was used to choose the optimal GE11 targeting density and evaluate the IC50 for free DOX, PEG-LP/DOX and GE11-LP/DOX. Qualitative and quantitative analysis of liposomes cellular uptake by A549 cells was detected by fluorescence microscope and flow cytometry. The real-time distribution of liposomes in A549 tumor-bearing nude micewas determined by a near-infrared (NIR) fluorescence imaging system. At last, the cellular uptake mechanism and the subcellular localization of the liposomes were examined by flow cytometry and confocal microscopy.Results: The results of 1H-NMR and FTIR verified the synthesis of theDSPE-PEG2000-GE11 and the calculated conjugated efficiency of GE11 to DSPE-PEG2000-Mal was above 90% detected by HPLC. The particle size of theAbstract Study of the anti-tum