Preparation Of Transferrin-modified Harmine Magnetoliposomes And Anti-brain Glioma Effects In Vitro And In Vivo

Objective:To prepare TF-HM-MPS with Fe₃O₄@CA as contrast agent and TF as targeting ligand,and to investigate its properties in vitro and in vivo.Methods:TF was used as a ligand,and it was attached to the surface of the magentoliposomes membrane by chemical coupling.TF-HM-MPS was prepared by reversed-phase sonication,and the most stable liposomes was selected by optimizing the prescription.In the study of anti-tumor cell activity in vitro,firstly,the toxicity of MPS to human glioma U87 cells was evaluated by cck-8.Secondly,the uptake of the liposomes by the cells was observed by FCM and LSCM.Finally,PI/AnnexinV staining,WB and PCR were used to investigate the effect of liposome on cell cycle and apoptosis of human glioma U87 cells.In the study of antitumor cell activity in vivo,firstly,the imaging effect of TF-MPS was observed by MRI.Secondly,the tumor-bearing mice were treated with TF-HM-MPS for21 days through the tail vein.Finally,the therapeutic effect was evaluated by calculating tumor sizes and observing pathological changes.Results:Fe₃O₄@CA was successfully prepared.The average particle sizes of DLS was?52.04±0.01?nm.The average size of Fe₃O₄@CA was about 10 nm by TEM.The content of Fe³⁺was 9.18 mmoL by ICP-MS,and it had strong T₂ effect,which showed it can be used as a contrast agent for MRI imaging.The prepared TF-HM-MPS was optimized.The optimal average particle sizes were?129.23±3.66?nm,the zeta potential was?-21.20±1.05?mV,and the encapsulation efficiency was?80.31±0.18?%,the drug loading efficiency was?1.01%±2.22?%.And TF-HM-MPS had good stability.With respect to the HM solution,TF-HM-MPS showed slower release rate and slower drug release rate,which were no significant difference from those results in PBS and plasma,TF-MPS also had a strong T₂ effect.The results of in vitro anti-tumor activity experiments showed that free HM was more toxic to human glioma U87,and the toxicity slowed down after HM was made into liposome.The targeting of TF to human glioma U87 cells was observed by FCM and LSCM.The results showed that TF-LPS was better than LPS.TF-HM-MPS was applied to human glioma U87 cells,and the apoptosis effect of TF-HM-MPS on human glioma U87 was verified by FCM,PCR and WB.The tumor-bearing mice were treated for 21 days.The results showed that TF-HM-LPS had obvious anti-tumor effect and less damage to tissues and organs.Conclusions:The experimental results show that the HM made TF-HM-MPS can significantly improve the therapeutic index of HM,and p53-mediated apoptosis and cycle pathway may be one of the principles of HM's anti-cancer mechanism,so TF-HM-MPS is an in vivo targeted drug delivery system with potential application in the treatment of cancer.This study can lay the foundation for the development of HM dosage form and its mechanism of anticancer action.